Sirtuin modulating compounds

ABSTRACT

Provided herein are novel sirtuin-modulating compounds and methods of use thereof. The sirtuin-modulating compounds may be used for increasing the lifespan of a cell, and treating and/or preventing a wide variety of diseases and disorders including, for example, diseases or disorders related to aging or stress, diabetes, obesity, neurodegenerative diseases, cardiovascular disease, blood clotting disorders, inflammation, cancer, and/or flushing as well as diseases or disorders that would benefit from increased mitochondrial activity. Also provided are compositions comprising a sirtuin-modulating compound in combination with another therapeutic agent.

RELATED APPLICATIONS

This application is a Continuation of U.S. patent application Ser. No.11/499,919, filed Aug. 4, 2006, which claims the benefit of U.S.Provisional Application Nos. 60/705,612, filed Aug. 4, 2005, 60/741,783,filed Dec. 2, 2005, 60/779,370, filed Mar. 3, 2006 and 60/792,276, filedApr. 14, 2006, the contents of which are incorporated by reference intheir entirety.

BACKGROUND

The Silent Information Regulator (SIR) family of genes represents ahighly conserved group of genes present in the genomes of organismsranging from archaebacteria to a variety of eukaryotes (Frye, 2000). Theencoded SIR proteins are involved in diverse processes from regulationof gene silencing to DNA repair. The proteins encoded by members of theSIR gene family show high sequence conservation in a 250 amino acid coredomain. A well-characterized gene in this family is S. cerevisiae SIR2,which is involved in silencing HM loci that contain informationspecifying yeast mating type, telomere position effects and cell aging(Guarente, 1999; Kaeberlein et al., 1999; Shore, 2000). The yeast Sir2protein belongs to a family of histone deacetylases (reviewed inGuarente, 2000; Shore, 2000). The Sir2 homolog, CobB, in Salmonellatyphimurium, functions as an NAD (nicotinamide adeninedinucleotide)-dependent ADP-ribosyl transferase (Tsang andEscalante-Semerena, 1998).

The Sir2 protein is a class III deacetylase which uses NAD as acosubstrate (Imai et al., 2000; Moazed, 2001; Smith et al., 2000; Tanneret al., 2000; Tanny and Moazed, 2001). Unlike other deacetylases, manyof which are involved in gene silencing, Sir2 is insensitive to class Iand II histone deacetylase inhibitors like trichostatin A (TSA) (Imai etal., 2000; Landry et al., 2000a; Smith et al., 2000).

Deacetylation of acetyl-lysine by Sir2 is tightly coupled to NADhydrolysis, producing nicotinamide and a novel acetyl-ADP ribosecompound (Tanner et al., 2000; Landry et al., 2000b; Tanny and Moazed,2001). The NAD-dependent deacetylase activity of Sir2 is essential forits functions which can connect its biological role with cellularmetabolism in yeast (Guarente, 2000; Imai et al., 2000; Lin et al.,2000; Smith et al., 2000). Mammalian Sir2 homologs have NAD-dependenthistone deacetylase activity (Imai et al., 2000; Smith et al., 2000).Most information about Sir2 mediated functions comes from the studies inyeast (Gartenberg, 2000; Gottschling, 2000).

Biochemical studies have shown that Sir2 can readily deacetylate theamino-terminal tails of histones H3 and H4, resulting in the formationof 1-O-acetyl-ADP -ribose and nicotinamide. Strains with additionalcopies of SIR2 display increased rDNA silencing and a 30% longer lifespan. It has recently been shown that additional copies of the C.elegans SIR2 homolog, sir-2.1, and the D. melanogaster dSir2 genegreatly extend life span in those organisms. This implies that theSIR2-dependent regulatory pathway for aging arose early in evolution andhas been well conserved. Today, Sir2 genes are believed to have evolvedto enhance an organism's health and stress resistance to increase itschance of surviving adversity.

SIRT3 is a homolog of SIRT1 that is conserved in prokaryotes andeukaryotes (P. Onyango et al., Proc. Natl. Acad. Sci. USA 99:13653-13658 (2002)). The SIRT3 protein is targeted to the mitochondrialcristae by a unique domain located at the N-terminus. SIRT3 hasNAD+-dependent protein deacetylase activity and is upbiquitouslyexpressed, particularly in metabolically active tissues. Upon transferto the mitochondria, SIRT3 is believed to be cleaved into a smaller,active form by a mitochondrial matrix processing peptidase (MPP) (B.Schwer et al., J. Cell Biol. 158: 647-657 (2002)).

Caloric restriction has been known for over 70 years to improve thehealth and extend the lifespan of mammals (Masoro, 2000). Yeast lifespan, like that of metazoans, is also extended by interventions thatresemble caloric restriction, such as low glucose. The discovery thatboth yeast and flies lacking the SIR2 gene do not live longer whencalorically restricted provides evidence that SIR2 genes mediate thebeneficial health effects of this diet (Anderson et al., 2003; Helfandand Rogina, 2004). Moreover, mutations that reduce the activity of theyeast glucose-responsive cAMP (adenosine 3′,5′-monophosphate)-dependent(PKA) pathway extend life span in wild type cells but not in mutant sir2strains, demonstrating that SIR2 is likely to be a key downstreamcomponent of the caloric restriction pathway (Lin et al., 2001).

SUMMARY

Provided herein are novel sirtuin-modulating compounds and methods ofuse thereof.

In one aspect, the invention provides sirtuin-modulating compounds ofFormula (I):

-   or a salt thereof, where:    -   Ring A is optionally substituted, fused to another ring or both;        and    -   Ring B is substituted with at least one carboxy, substituted or        unsubstituted arylcarboxamine, substituted or unsubstituted        aralkylcarboxamine, substituted or unsubstituted heteroaryl        group, substituted or unsubstituted heterocyclylcarbonylethenyl,        or polycyclic aryl group or is fused to an aryl ring and is        optionally substituted by one or more additional groups.

In another aspect, the invention provides sirtuin-modulating compoundsof Formula (II):

-   or a salt thereof, where:    -   Ring A is optionally substituted;    -   R₁, R₂, R₃ and R₄ are independently selected from the group        consisting of —H, halogen, —OR₅, —CN, —CO₂R₅, —OCOR₅, —OCO₂R₅,        —C(O)NR₅R₆, —OC(O)NR₅R₆, —C(O)R₅, —COR₅, —SR₅, —OSO₃H,        —S(O)_(n)R₅, —S(O)_(n)OR₅, —S(O)_(n)NR₅R₆, —NR₅R₆, —NR₅C(O)OR₆,        —NR₅C(O)R₆ and —NO₂;    -   R₅ and R₆ are independently —H, a substituted or unsubstituted        alkyl group, a substituted or unsubstituted aryl group or a        substituted or unsubstituted heterocyclic group; and    -   n is 1 or 2.

In yet another aspect, the invention provides sirtuin-modulatingcompounds of Formula (III):

-   or a salt thereof, where:    -   Ring A is optionally substituted;    -   R₅ and R₆ are independently —H, a substituted or unsubstituted        alkyl group, a substituted or unsubstituted aryl group or a        substituted or unsubstituted heterocyclic group;    -   R₇, R₉, R₁₀ and R₁₁ are independently selected from the group        consisting of —H, halogen, —R₅, —OR₅, —CN, —CO₂R₅, —OCOR₅,        —OCO₂R₅, —C(O)NR₅R₆, —OC(O)NR₅R₆, —C(O)R₅, —COR₅, —SR₅, —OSO₃H,        —S(O)_(n)R₅, —S(O)_(n)OR₅, —S(O)_(n)NR₅R₆, —NR₅R₆, —NR₅C(O)OR₆,        —NR₅C(O)R₆ and —NO₂;    -   R₈ is a polycyclic aryl group; and    -   n is 1 or 2.

In another aspect, the invention provides sirtuin-modulating compoundsof Formula (IV):Ar-L-J-M-K—Ar′  (IV)

-   or a salt thereof, wherein:    -   each Ar and Ar′ is independently an optionally substituted        carbocyclic or heterocyclic aryl group;    -   L is an optionally substituted carbocyclic or heterocyclic        arylene group;    -   each J and K is independently NR₁′, O, S, or is optionally        independently absent; or when J is NR₁′, R₁′ is a C1-C4 alkylene        or C2-C4 alkenylene attached to Ar′ to form a ring fused to Ar′;        or when K is NR₁′, R₁′ is a C1-C4 alkylene or C2-C4 alkenylene        attached to L to form a ring fused to L;    -   each M is C(O), S(O), S(O)₂, or CR₁′R₁′;    -   each R₁′ is independently selected from H, C1-C10 alkyl; C2-C10        alkenyl; C2-C10 alkynyl; C3-C10 cycloalkyl; C4-C10 cycloalkenyl;        aryl; R₅′; halo; haloalkyl; CF₃; SR₂′; OR₂′; NR₂′R₂′; NR₂′R₃′;        COOR₂′; NO₂; CN; C(O)R₂′; C(O)C(O)R₂′; C(O)NR₂′R₂′; OC(O)R₂′;        S(O)₂R₂′; S(O)₂NR₂′R₂′; NR₂′C(O)NR₂′R₂′; NR₂′C(O)C(O)R₂′;        NR₂′C(O)R₂′; NR₂′(COOR₂′); NR₂′C(O)R₅′; NR₂′S(O)₂NR₂′R₂′;        NR₂′S(O)₂R₂′; NR₂′S(O)₂R₅′; NR₂′C(O)C(O)NR₂′R₂′;        NR₂′C(O)C(O)NR₂′R₃′; C1-C10 alkyl substituted with aryl, R₄′ or        R₅′; or C2-C10 alkenyl substituted with aryl, R₄′ or R₅′;    -   each R₂′ is independently H; C1-C10 alkyl; C2-C10 alkenyl;        C2-C10 alkynyl; C3-C10 cycloalkyl; C4-C10 cycloalkenyl; aryl;        R₆′; C1-C10 alkyl substituted with 1-3 independent aryl, R₄′ or        R₆′ groups; C3-C10 cycloalkyl substituted with 1-3 independent        aryl, R₄′ or R₆′ groups; or C2-C10 alkenyl substituted with 1-3        independent aryl, R₄′ or R₆′;    -   each R₃′ is independently C(O)R₂′, COOR₂′, or S(O)₂R₂′;    -   each R₄′ is independently halo, CF₃, SR₇′, OR₇′, OC(O)R₇′,        NR₇′R₇′, NR₇′R₈′, NR₈′R₈′, COOR₇′, NO₂, CN, C(O)R₇′, or        C(O)NR₇′R₇′;    -   each R₅′ is independently a 5-8 membered monocyclic, 8-12        membered bicyclic, or 11-14 membered tricyclic ring system        comprising 1-3 heteroatoms if monocyclic, 1-6 heteroatoms if        bicyclic, or 1-9 heteroatoms if tricyclic, said heteroatoms        selected from O, N, or S, which may be saturated or unsaturated,        and wherein 0, 1, 2 or 3 atoms of each ring may be substituted        by a substituent independently selected from C1-C10 alkyl;        C2-C10 alkenyl; C2-C10 alkynyl; C3-C10 cycloalkyl; C4-C10        cycloalkenyl; aryl; R₆′; halo; sulfur; oxygen; CF₃; haloalkyl;        SR₂′; OR₂′; OC(O)R₂′; NR₂′R₂′; NR₂′R₃′; NR₃′R₃′; COOR₂′; NO₂;        CN; C(O)R₂′; C(O)NR₂′R₂′; C1-C10 alkyl substituted with 1-3        independent R₄′, R₆′, or aryl; or C2-C10 alkenyl substituted        with 1-3 independent R₄′, R₆′, or aryl;    -   each R₆′ is independently a 5-8 membered monocyclic, 8-12        membered bicyclic, or 11-14 membered tricyclic ring system        comprising 1-3 heteroatoms if monocyclic, 1-6 heteroatoms if        bicyclic, or 1-9 heteroatoms if tricyclic, said heteroatoms        selected from O, N, or S, which may be saturated or unsaturated,        and wherein 0, 1, 2 or 3 atoms of each ring may be substituted        by a substituent independently selected from C1-C10 alkyl;        C2-C10 alkenyl; C2-C10 alkynyl; C3-C10 cycloalkyl; C4-C10        cycloalkenyl; halo; sulfur; oxygen; CF₃; haloalkyl; SR₇′; OR₇′;        NR₇′R₇′; NR₇′R₈′; NR₈′R₈′; COOR₇′; NO₂; CN; C(O)R₇′; or        C(O)NR₇′R₇′;    -   each R₇′ is independently H, C1-C10 alkyl; C2-C10 alkenyl;        C2-C10 alkynyl; C3-C10 cycloalkyl; C4-C10 cycloalkenyl;        haloalkyl; C1-C10 alkyl optionally substituted with 1-3        independent C1-C10 alkyl, C2-C10 alkenyl, C2-C10 alkynyl, C3-C10        cycloalkyl, C4-C10 cycloalkenyl, halo, CF₃, OR₁₀′, SR₁₀′,        NR₁₀′R₁₀′, COOR₁₀′, NO₂, CN, C(O)R₁₀′, C(O)NR₁₀′R₁₀′,        NHC(O)R₁₀′, or OC(O)R₁₀′; or phenyl optionally substituted with        1-3 independent C1-C10 alkyl, C2-C10 alkenyl, C2-C10 alkynyl,        C3-C10 cycloalkyl, C4-C10 cycloalkenyl, halo, CF₃, OR₁₀′, SR₁₀′,        NR₁₀′R₁₀′, COOR₁₀′, NO₂, CN, C(O)R₁₀′, C(O)NR₁₀′R₁₀′,        NHC(O)R₁₀′, or OC(O)R₁₀;    -   each R₈′ is independently C(O)R₇′, COOR₇′, or S(O)₂R₇′;    -   each R₉′ is independently H, C1-C10 alkyl, C2-C10 alkenyl,        C2-C10 alkynyl, C3-C10 cycloalkyl, C4-C10 cycloalkenyl, or        phenyl optionally substituted with 1-3 independent C1-C10 alkyl,        C2-C10 alkenyl, C2-C10 alkynyl, C3-C10 cycloalkyl, C4-C10        cycloalkenyl, halo, CF₃, OR₁₀′, SR₁₀′, NR₁₀′R₁₀′, COOR₁₀′, NO₂,        CN, C(O)R₁₀′, C(O)NR₁₀′R₁₀′, NHC(O)R₁₀′, or OC(O)R₁₀′;    -   each R₁₀′ is independently H; C1-C10 alkyl; C2-C10 alkenyl;        C2-C10 alkynyl; C3-C10 cycloalkyl; C4-C10 cycloalkenyl; C1-C10        alkyl optionally substituted with halo, CF₃, OR₁₁′, SR₁₁′,        NR₁₁′R₁₁′, COOR₁₁′, NO₂, CN; or phenyl optionally substituted        with halo, CF₃, OR₁₁′, SR₁₁′, NR₁₁′R₁₁′, COOR₁₁′, NO₂, CN;    -   each R₁₁′ is independently H; C1-C10 alkyl; C3-C10 cycloalkyl or        phenyl;    -   each haloalkyl is independently a C1-C10 alkyl substituted with        one or more halogen atoms, selected from F, Cl, Br, or I,        wherein the number of halogen atoms may not exceed that number        that results in a perhaloalkyl group; and    -   each aryl is independently optionally substituted with 1-3        independent C1-C10 alkyl; C2-C10 alkenyl; C2-C10 alkynyl; C3-C10        cycloalkyl; C4-C10 cycloalkenyl; R₆′; halo; haloalkyl; CF₃;        OR₉′; SR₉′; NR₉′R₉′; COOR₉′; NO₂; CN; C(O)R₉′; C(O)C(O)R₉′;        C(O)NR₉′R₉′; S(O)₂R₉′; N(R₉′)C(O)R₉′; N(R₉′)(COOR₉′);        N(R₉′)S(O)₂R₉′; S(O)₂NR₉′R₉′; OC(O)R₉′; NR₉′C(O)NR₉′R₉′;        NR₉′C(O)C(O)R₉′; NR₉′C(O)R₆′; NR₉′S(O)₂NR₉′R₉′; NR₉′S(O)₂R₆′;        NR₉′C(O)C(O)NR₉′R₉′; C1-C10 alkyl substituted with 1-3        independent R₆′, halo, CF₃, OR₉′, SR₉′, NR₉′R₉′, COOR₉′, NO₂,        CN, C(O)R₉′, C(O)NR₉′R₉′, NHC(O)R₉′, NH(COOR₉′), S(O)₂NR₉′R₉′,        OC(O)R₉′; C2-C10 alkenyl substituted with 1-3 independent R₆′,        halo, CF₃, OR₉′, SR₉′, NR₉′R₉′, COOR₉′, NO₂, CN, C(O)R₉′,        C(O)NR₉′R₉′, NHC(O)R₉′, NH(COOR₉′), S(O)₂NR₉′R₉′, OC(O)R₉′; or        R₉′.

In a further aspect, the invention provides sirtuin-modulating compoundsof Formula (IVa):Het-L-Q-Ar′  (IVa)

-   or a salt thereof, where:    -   Het is an optionally substituted heterocyclic aryl group;    -   L is an optionally substituted carbocyclic or heterocyclic        arylene group;    -   Ar′ is an optionally substituted carbocyclic or heterocyclic        aryl group; and    -   Q is selected from —NR₁′—C(O)—, —NR₁′—S(O)₂—, —NR₁′—C(O)—NR₁′—,        —NR₁′—C(S)—NR₁′—, —NR₁′—C(O)—CR₁′R₁′—NR₁′—,        —NR₁′—C(═NR₁′)—NR₁′—, —C(O)—NR₁′—, —C(O)—NR₁′—S(O)₂—, —NR₁′—,        —CR₁′R₁′—, —NR₁′—C(O)—CR₁′═CR₁′—, —NR₁′—S(O)₂—NR₁′—,        —NR₁′—C(O)—NR₁′—S(O)₂—, —NR₁′—CR₁′R₁′—C(O)—NR₁′—,        —CR₁′R₁′—C(O)—NR₁′—, —NR₁′—C(O)—CR₁′═CR₁′—CR₁′R₁′—,        —NR₁′—C(═N—CN)—NR₁′—, —NR₁′—C(O)—CR₁′R₁′—,

-   -   each R₁′ is independently selected from H or optionally        substituted C₁-C₃ straight or branched alkyl, wherein:

-   when Het is a polycyclic heteroaryl, L is an optionally substituted    phenylene, Q and Het are attached to L in a meta orientation, and    Ar′ is optionally substituted phenyl; then Q is not —NH—C(O)—.

In still yet another aspect, the invention provides sirtuin-modulatingcompounds of Formula (V):

-   -   or a salt thereof, wherein:    -   Ring A is optionally substituted with at least one R₁′ group;    -   Y₁, Y₂, Y₃, Y₄, and Y₅ are independently R₁′;    -   each R₁′ is independently selected from H, C1-C10 alkyl; C2-C10        alkenyl; C2-C10 alkynyl; C3-C10 cycloalkyl; C4-C10 cycloalkenyl;        aryl; R₅′; halo; haloalkyl; CF₃; SR₂′; OR₂′; NR₂′R₂′; NR₂′R₃′;        COOR₂′; NO₂; CN; C(O)R₂′; C(O)C(O)R₂′; C(O)NR₂′R₂′; OC(O)R₂′;        S(O)₂R₂′; S(O)₂NR₂′R₂′; NR₂′C(O)NR₂′R₂′; NR₂′C(O)C(O)R₂′;        NR₂′C(O)R₂′; NR₂′(COOR₂′); NR₂′C(O)R₅′; NR₂′S(O)₂NR₂′R₂′;        NR₂′S(O)₂R₂′; NR₂′S(O)₂R₅′; NR₂′C(O)C(O)NR₂′R₂′;        NR₂′C(O)C(O)NR₂′R₃′; C1-C10 alkyl substituted with aryl, R₄′ or        R₅′; or C2-C10 alkenyl substituted with aryl, R₄′ or R₅′;    -   each R₂′ is independently H; C1-C10 alkyl; C2-C10 alkenyl;        C2-C10 alkynyl; C3-C10 cycloalkyl; C4-C10 cycloalkenyl; aryl;        R₆′; C1-C10 alkyl substituted with 1-3 independent aryl, R₄′ or        R₆′ groups; C3-C10 cycloalkyl substituted with 1-3 independent        aryl, R₄′ or R₆′ groups; or C2-C10 alkenyl substituted with 1-3        independent aryl, R₄′ or R₆′;    -   each R₃′ is independently C(O)R₂′, COOR₂′, or S(O)₂R₂′;    -   each R₄′ is independently halo, CF₃, SR₇′, OR₇′, OC(O)R₇′,        NR₇′R₇′, NR₇′R₈′, NR₈′R₈′, COOR₇′, NO₂, CN, C(O)R₇′, or        C(O)NR₇′R₇′;    -   each R₅′ is independently a 5-8 membered monocyclic, 8-12        membered bicyclic, or 11-14 membered tricyclic ring system        comprising 1-3 heteroatoms if monocyclic, 1-6 heteroatoms if        bicyclic, or 1-9 heteroatoms if tricyclic, said heteroatoms        selected from O, N, or S, which may be saturated or unsaturated,        and wherein 0, 1, 2 or 3 atoms of each ring may be substituted        by a substituent independently selected from C1-C10 alkyl;        C2-C10 alkenyl; C2-C10 alkynyl; C3-C10 cycloalkyl; C4-C10        cycloalkenyl; aryl; R₆′; halo; sulfur; oxygen; CF₃; haloalkyl;        SR₂′; OR₂′; OC(O)R₂′; NR₂′R₂′; NR₂′R₃′; NR₃′R₃′; COOR₂′; NO₂;        CN; C(O)R₂′; C(O)NR₂′R₂′; C1-C10 alkyl substituted with 1-3        independent R₄′, R₆′, or aryl; or C2-C10 alkenyl substituted        with 1-3 independent R₄′, R₆′, or aryl;    -   each R₆′ is independently a 5-8 membered monocyclic, 8-12        membered bicyclic, or 11-14 membered tricyclic ring system        comprising 1-3 heteroatoms if monocyclic, 1-6 heteroatoms if        bicyclic, or 1-9 heteroatoms if tricyclic, said heteroatoms        selected from O, N, or S, which may be saturated or unsaturated,        and wherein 0, 1, 2 or 3 atoms of each ring may be substituted        by a substituent independently selected from C1-C10 alkyl;        C2-C10 alkenyl; C2-C10 alkynyl; C3-C10 cycloalkyl; C4-C10        cycloalkenyl; halo; sulfur; oxygen; CF₃; haloalkyl; SR₇′; OR₇′;        NR₇′R₇′; NR₇′R₈′; NR₈′R₈′; COOR₇′; NO₂; CN; C(O)R₇′; or        C(O)NR₇′R₇′;    -   each R₇′ is independently H, C1-C10 alkyl; C2-C10 alkenyl;        C2-C10 alkynyl; C3-C10 cycloalkyl; C4-C10 cycloalkenyl;        haloalkyl; C1-C10 alkyl optionally substituted with 1-3        independent C1-C10 alkyl, C2-C10 alkenyl, C2-C10 alkynyl, C3-C10        cycloalkyl, C4-C10 cycloalkenyl, halo, CF₃, OR₁₀′, SR₁₀′,        NR₁₀′R₁₀′, COOR₁₀′, NO₂, CN, C(O)R₁₀′, C(O)NR₁₀′R₁₀′,        NHC(O)R₁₀′, or OC(O)R₁₀′; or phenyl optionally substituted with        1-3 independent C1-C10 alkyl, C2-C10 alkenyl, C2-C10 alkynyl,        C3-C10 cycloalkyl, C4-C10 cycloalkenyl, halo, CF₃, OR₁₀′, SR₁₀′,        NR₁₀′R₁₀′, COOR₁₀′, NO₂, CN, C(O)R₁₀′, C(O)NR₁₀′R₁₀′,        NHC(O)R₁₀′, or OC(O)R₁₀′;    -   each R₈′ is independently C(O)R₇′, COOR₇′, or S(O)₂R₇′;    -   each R₉′ is independently H, C1-C10 alkyl, C2-C10 alkenyl,        C2-C10 alkynyl, C3-C10 cycloalkyl, C4-C10 cycloalkenyl, or        phenyl optionally substituted with 1-3 independent C1-C10 alkyl,        C2-C10 alkenyl, C2-C10 alkynyl, C3-C10 cycloalkyl, C4-C10        cycloalkenyl, halo, CF₃, OR₁₀′, SR₁₀′, NR₁₀′R₁₀′, COOR₁₀′, NO₂,        CN, C(O)R₁₀′, C(O)NR₁₀′R₁₀′, NHC(O)R₁₀′, or OC(O)R₁₀′;    -   each R₁₀′ is independently H; C1-C10 alkyl; C2-C10 alkenyl;        C2-C10 alkynyl; C3-C10 cycloalkyl; C4-C10 cycloalkenyl; C1-C10        alkyl optionally substituted with halo, CF₃, OR₁₁′, SR₁₁′,        NR₁₁′R₁₁′, COOR₁₁′, NO₂, CN; or phenyl optionally substituted        with halo, CF₃, OR₁₁′, SR₁₁′, NR₁₁′R₁₁′, COOR₁₁′, NO₂, CN;    -   each R₁₁′ is independently H; C1-C10 alkyl; C3-C10 cycloalkyl or        phenyl;    -   each haloalkyl is independently a C1-C10 alkyl substituted with        one or more halogen atoms, selected from F, Cl, Br, or I,        wherein the number of halogen atoms may not exceed that number        that results in a perhaloalkyl group; and    -   each aryl is independently a 5-to 7-membered monocyclic ring        system or a 9-to 12-membered bicyclic ring system optionally        substituted with 1-3 independent C1-C10 alkyl; C2-C10 alkenyl;        C2-C10 alkynyl; C3-C10 cycloalkyl; C4-C10 cycloalkenyl; R₆′;        halo; haloalkyl; CF₃; OR₉′; SR₉′; NR₉′R₉′; COOR₉′; NO₂; CN;        C(O)R₉′; C(O)C(O)R₉′; C(O)NR₉′R₉′; S(O)₂R₉′; N(R₉′)C(O)R₉′;        N(R₉′)(COOR₉′); N(R₉′)S(O)₂R₉′; S(O)₂NR₉′R₉′; OC(O)R₉′;        NR₉′C(O)NR₉′R₉′; NR₉′C(O)C(O)R₉′; NR₉′C(O)R₆′; NR₉′S(O)₂NR₉′R₉′;        NR₉′S(O)₂R₆′; NR₉′C(O)C(O)NR₉′R₉′; C1-C10 alkyl substituted with        1-3 independent R₆′, halo, CF₃, OR₉′, SR₉′, NR₉′R₉′, COOR₉′,        NO₂, CN, C(O)R₉′, C(O)NR₉′R₉′, NHC(O)R₉′, NH(COOR₉′),        S(O)₂NR₉′R₉′, OC(O)R₉′; C2-C10 alkenyl substituted with 1-3        independent R₆′, halo, CF₃, OR₉′, SR₉′, NR₉′R₉′, COOR₉′, NO₂,        CN, C(O)R₉′, C(O)NR₉′R₉′, NHC(O)R₉′, NH(COOR₉′), S(O)₂NR₉′R₉′,        OC(O)R₉′; or R₉′.

In one aspect, the invention provides sirtuin-modulating compounds ofStructural Formula (VI):

-   or a salt thereof, wherein:    -   Het is an optionally substituted heterocyclic aryl group; and    -   Ar′ is an optionally substituted carbocyclic or heterocyclic        aryl group.

The invention also includes prodrugs and metabolites of the compoundsdisclosed herein.

In another aspect, the invention provides sirtuin-modulating compoundsof Structural Formula (VII):

-   or a salt thereof, wherein:    -   each of X₇, X₈, X₉ and X₁₀ is independently selected from N,        CR²⁰, or CR₁′, wherein:        -   each R²⁰ is independently selected from H or a solubilizing            group;        -   each R₁′ is independently selected from H or optionally            substituted C₁-C₃ straight or branched alkyl;        -   one of X₇, X₈, X₉ and X₁₀ is N and the others are selected            from CR₁′; and        -   zero to one R²⁰ is a solubilizing group;    -   R¹⁹ is selected from:

-   -   wherein:        -   each Z₁₀, Z₁₁, Z₁₂ and Z₁₃ is independently selected from N,            CR²⁰, or CR₁′; and        -   each Z₁₄, Z₁₅ and Z₁₆ is independently selected from N,            NR₁′, S, O, CR²⁰, or CR₁′, wherein:        -   zero to two of Z₁₀, Z₁₁, Z₁₂ or Z₁₃ are N;        -   at least one of Z₁₄, Z₁₅ and Z₁₆ is N, NR₁′, S or O;        -   zero to one of Z₁₄, Z₁₅ and Z₁₆ is S or O;        -   zero to two of Z₁₄, Z₁₅ and Z₁₆ are N or NR₁′;        -   zero to one R²⁰ is a solubilizing group;        -   zero to one R₁′ is an optionally substituted C₁-C₃ straight            or branched alkyl; and    -   R²¹ is selected from —NR₁′—C(O)—, —NR₁′—S(O)₂—,        —NR₁′—C(O)—NR₁′—, —NR₁′—C(S)—NR₁′—, —NR₁′—C(S)—NR₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—NR₁′—, —NR₁′—C(═NR₁′)—NR₁′—, —C(O)—NR₁′—,        —C(O)—NR₁′—S(O)₂—, —NR₁′—, —CR₁′R₁′—, —NR₁′—C(O)—CR₁′═CR₁′—,        —NR₁′—S(O)₂—NR₁′—, —NR₁′—C(O)—NR₁′—S(O)₂—,        —NR₁′—CR₁′R₁′—C(O)—NR₁′—, —CR₁′R₁′—C(O)—NR₁′—,        —NR₁′—C(O)—CR₁′═CR₁′—CR₁′R₁′—, —NR₁′—C(═N—CN)—NR₁′—,        —NR₁′—C(O)—CR₁′R₁′—O—, —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—O—,        —NR₁′—S(O)₂—CR₁′R₁′—, —NR₁′—S(O)₂—CR₁′R₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—; —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—,        —NR₁′—C(S)—NR₁′—CR₁′R₁′—CR₁′R₁′—, —NR₁′—C(O)—O—,

-   -   R³¹ is selected from an optionally substituted monocyclic or        bicyclic aryl, or an optionally substituted monocyclic or        bicyclic heteroaryl, with the provisos that said compound is        not:

-   -    that when R¹⁹ is

-   -    and R²¹ is —NHC(O)—, R³¹ is not an optionally substituted        phenyl.

In certain embodiments, compounds of Structural Formula (VII) have thefollowing values:

-   -   each of X₇, X₈, X₉ and X₁₀ is independently selected from N,        CR²⁰, or CR₁′, wherein:    -   each R²⁰ is independently selected from H or a solubilizing        group;    -   each R₁′ is independently selected from H or optionally        substituted C₁-C₃ straight or branched alkyl;    -   one of X₇, X₈, X₉ and X₁₀ is N and the others are selected from        CR²⁰ or CR₁′; and    -   zero to one R²⁰ is a solubilizing group;    -   R¹⁹ is selected from:

-   -   wherein:        -   each Z₁₀, Z₁₀, Z₁₂ and Z₁₃ is independently selected from N,            CR²⁰, or CR₁′; and        -   each Z₁₄, Z₁₅ and Z₁₆ is independently selected from N,            NR₁′, S, O, CR²⁰, or CR₁′, wherein:        -   zero to two of Z₁₀, Z₁₁, Z₁₂ or Z₁₃ are N;        -   at least one of Z₁₄, Z₁₅ and Z₁₆ is N, NR₁′, S or O;        -   zero to one of Z₁₄, Z₁₅ and Z₁₆ is S or O;        -   zero to two of Z₁₄, Z₁₅ and Z₁₆ are N or NR₁′;        -   zero to one R²⁰ is a solubilizing group;        -   zero to one R₁′ is an optionally substituted C₁-C₃ straight            or branched alkyl; and    -   R²¹ is selected from —NR₁′—C(O)—, —NR₁′—S(O)₂—,        —NR₁′—C(O)—NR₁′—, —NR₁′—C(S)—NR₁′—, —NR₁′—C(S)—NR₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—NR₁′—, —NR₁′—C(═NR₁′)—NR₁′—, —C(O)—NR₁′—,        —C(O)—NR₁′—S(O)₂—, —NR₁′—, —CR₁′R₁′—, —NR₁′—C(O)—CR₁′═CR₁′—,        —NR₁′—S(O)₂—NR₁′—, —NR₁′—C(O)—NR₁′—S(O)₂—,        —NR₁′—CR₁′R₁′—C(O)—NR₁′—, —CR₁′R₁′—C(O)—NR₁′—,        —NR₁′—C(O)—CR₁′═CR₁′—CR₁′R₁′—, —NR₁′—C(═N—CN)—NR₁′—,        —NR₁′—C(O)—CR₁′R₁′—O—, —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—O—,        —NR₁′—S(O)₂—CR₁′R₁′—, —NR₁′—S(O)₂—CR₁′R₁′—CR₁′R₁′—, or        —NR₁′—C(O)—CR₁′R₁′—; and    -   R³¹ is selected from an optionally substituted monocyclic or        bicyclic aryl, or an optionally substituted monocyclic or        bicyclic heteroaryl, with the provisos that:    -   said compound is not:

-   -   when X₈ and X₉ are each independently selected from CR²⁰ or        CR₁′, R¹⁹ is

-   -    and each of Z₁₀, Z₁₁, Z₁₂ and Z₁₃ is independently selected        from CR²⁰, or CR₁′, then:        -   a) at least one of X₈ and X₉ is not CH; or        -   b) at least one of Z₁₀, Z₁₁, Z₁₂ and Z₁₃ is CR²⁰, wherein            R²⁰ is a solubilizing group.

In yet another embodiment, the invention provides sirtuin-modulatingcompounds of Structural Formula (VIII):

-   or a salt thereof, wherein:    -   R₁′ is selected from H or optionally substituted C₁-C₃ straight        or branched alkyl;    -   R²¹ is selected from —NR₁′—C(O)—, —NR₁′—S(O)₂—,        —NR₁′—C(O)—NR₁′—, —NR₁′—C(S)—NR₁′—, —NR₁′—C(S)—NR₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—NR₁′—, —NR₁′—C(═NR₁′)—NR₁′—, —C(O)—NR₁′—,        —C(O)—NR₁′—S(O)₂—, —NR₁′—, —CR₁′R₁′—, —NR₁′—C(O)—CR₁′═CR₁′—,        —NR₁′—S(O)₂—NR₁′—, —NR₁′—C(O)—NR₁′—S(O)₂—,        —NR₁′—CR₁′R₁′—C(O)—NR₁′—, —CR₁′R₁′—C(O)—NR₁′—,        —NR₁′—C(O)—CR₁′═CR₁′—CR₁′R₁′—, —NR₁′—C(═N—CN)—NR₁′—,        —NR₁′—C(O)—CR₁′R₁′—O—, —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—O—,        —NR₁′—S(O)₂—CR₁′R₁′—, —NR₁′—S(O)₂—CR₁′R₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—; —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—,        —NR₁′—C(S)—NR₁′—CR₁′R₁′—CR₁′R₁′—, —NR₁′—C(O)—O—,

-   -   R³¹ is selected from an optionally substituted monocyclic or        bicyclic aryl, or an optionally substituted monocyclic or        bicyclic heteroaryl, with the provisos that:    -   when R₁′ is methyl, and R²¹ is —NH—C(O)—, R³¹ is not

-   -    1-methoxynaphthyl; 2-methoxynaphthyl; or unsubstituted        2-thienyl;    -   when R₁′ is methyl, and R²¹ is —NH—C(O)—CH═CH—, R³¹ is not

-   -   when R₁′ is methyl, and R²¹ is —NH—C(O)—CH—O—, R³¹ is not        unsubstituted naphthyl; 2-methoxy, 4-nitrophenyl; 4-chloro,        2-methylphenyl; or 4-t-butylphenyl; and    -   when R²¹ is —NH—C(O)—, R³¹ is not optionally substituted phenyl.

In a further embodiment, the invention provides sirtuin-modulatingcompounds of Structural Formula (IX):

-   or a salt thereof, wherein:    -   R₁′ is selected from H or optionally substituted C₁-C₃ straight        or branched alkyl; and    -   R⁵⁰ is selected from 2,3-dimethoxyphenyl, phenoxyphenyl,        2-methyl-3-methoxyphenyl, 2-methoxy-4-methylphenyl, or phenyl        substituted with 1 to 3 substituents, wherein one of said        substituents is a solubilizing group; with the provisos that R⁵⁰        is not substituted simultaneously with a solubilizing group and        a nitro group, and R⁵⁰ is not singly substituted at the        4-position with cyclic solubilizing group or at the 2-position        with a morpholino group.

In one aspect, the invention provides sirtuin-modulating compounds ofStructural Formula (X):

-   or a salt thereof, wherein:    -   R₁′ is selected from H or optionally substituted C₁-C₃ straight        or branched alkyl; and    -   R⁵¹ is selected from an optionally substituted monocyclic        heteroaryl, an optionally substituted bicyclic heteroaryl, or an        optionally substituted naphthyl, wherein R⁵¹ is not        chloro-benzo[b]thienyl, unsubstituted benzodioxolyl,        unsubstituted benzofuranyl, methyl-benzofuranyl, unsubstituted        furanyl, phenyl-, bromo-, or nitro-furyl,        chlorophenyl-isoxazolyl, oxobenzopyranyl, unsubstituted        naphthyl, methoxy-, methyl-, or halo-naphthyl, unsubstituted        thienyl, unsubstituted pyridinyl, or chloropyridinyl.

In another aspect, the invention provides sirtuin-modulating compoundsof Structural Formula (XI):

-   or a salt thereof, wherein:    -   R₁′ is selected from H or optionally substituted C₁-C₃ straight        or branched alkyl;    -   R²² is selected from —NR²³—C(O)—, —NR₁′—S(O)₂—,        —NR₁′—C(O)—NR₁′—, —NR₁′—C(S)—NR₁′—, —NR₁′—C(S)—NR₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—NR₁′—, —NR₁′—C(═NR₁′)—NR₁′—, —C(O)—NR₁′—,        —C(O)—NR₁′—S(O)₂—, —NR₁′—, —CR₁′R₁′—, —NR₁′—C(O)—CR₁′═CR₁′—,        —NR₁′—S(O)₂—NR₁′—, —NR₁′—C(O)—NR₁′—S(O)₂—,        —NR₁′—CR₁′R₁′—C(O)—NR₁′—, —CR₁′R₁′—C(O)—NR₁′—,        —NR₁′—C(O)—CR₁′═CR₁′—CR₁′R₁′—, —NR₁′—C(═N—CN)—NR₁′—,        —NR₁′—C(O)—CR₁′R₁′—O—, —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—O—,        —NR₁′—S(O)₂—CR₁′R₁′—, —NR₁′—S(O)₂—CR₁′R₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—, —NR₁′—C(S)—NR₁′—CR₁′R₁′—CR₁′R₁′—,        —NR₁′—C(O)—O— or —NR₁′—C(O)—CR₁′R₁′—, wherein R²³ is an        optionally substituted C₁-C₃ straight or branched alkyl; and    -   R³¹ is selected from an optionally substituted monocyclic or        bicyclic aryl, or an optionally substituted monocyclic or        bicyclic heteroaryl, with the provisos that:    -   when R²² is —NH—C(O)—CH═CH—, R³¹ is not unsubstituted furyl,        5-(2-methyl-3-chlorophenyl)-furanyl, 2,4-dichlorophenyl,        3,5-dichloro-2-methoxyphenyl, 3-nitrophenyl, 4-chlorophenyl,        4-chloro-3-nitrophenyl, 4-isopropylphenyl, 4-methoxyphenyl,        2-methoxy-5-bromophenyl, or unsubstituted phenyl;    -   when R²² is —NH—C(O)—CH₂—, R³¹ is not 3,4-dimethoxyphenyl,        4-chlorophenyl, or unsubstituted phenyl;    -   when R²² is —NH—C(O)—CH₂—O—, R³¹ is not        2,4-dimethyl-6-nitrophenyl, 2-or 4-nitrophenyl,        4-cyclohexylphenyl, 4-methoxyphenyl, unsubstituted naphthyl, or        unsubstituted phenyl, or phenyl monosubstituted, disubstituted        or trisubstituted solely with substituents selected from        straight-or branched-chain alkyl or halo;    -   when R²² is —NH—C(O)—CH(CH₃)—O—, R³¹ is not 2,4-dichlorophenyl,        4-chlorophenyl, or unsubstituted phenyl; and    -   when R²² is —NH—S(O)₂—, R³¹ is not unsubstituted phenyl.

In yet another aspect, the invention provides sirtuin-modulatingcompounds of Structural Formula (XII):

-   or a salt thereof, wherein:    -   each of X₇, X₈, X₉ and X₁₀ is independently selected from N,        CR²⁰, or CR₁′, wherein:        -   each R²⁰ is independently selected from H or a solubilizing            group;        -   each R₁′ is independently selected from H or optionally            substituted C₁-C₃ straight or branched alkyl;        -   one of X₇, X₈, X₉ and X₁₀ is N and the others are selected            from CR²⁰ or CR₁′; and        -   zero to one R²⁰ is a solubilizing group;    -   R¹⁹ is selected from:

-   -   wherein:        -   each Z₁₀, Z₁₁, Z₁₂ and Z₁₃ is independently selected from N,            CR²⁰, or CR₁′; and        -   each Z₁₄, Z₁₅ and Z₁₆ is independently selected from N,            NR₁′, S, O, CR²⁰, or CR₁′, wherein:        -   zero to two of Z₁₀, Z₁₁, Z₁₂ or Z₁₃ are N;        -   at least one of Z₁₄, Z₁₅ and Z₁₆ is N, NR₁′, O or S;        -   zero to one of Z₁₄, Z₁₅ and Z₁₆ is S or O;        -   zero to two of Z₁₄, Z₁₅ and Z₁₆ are N or NR₁′;        -   zero to one R²⁰ is a solubilizing group;        -   zero to one R₁′ is an optionally substituted C₁-C₃ straight            or branched alkyl; and    -   R²¹ is selected from —NR₁′—C(O)—, —NR₁′—S(O)₂—,        —NR₁′—C(O)—NR₁′—, —NR₁′—C(S)—NR₁′—, —NR₁′—C(S)—NR₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—NR₁′—, —NR₁′—C(═NR₁′)—NR₁′—, —C(O)—NR₁′—,        —C(O)—NR₁′—S(O)₂—, —NR₁′—, —CR₁′R₁′—, —NR₁′—C(O)—CR₁′═CR₁′—,        —NR₁′—S(O)₂—NR₁′—, —NR₁′—C(O)—NR₁′—S(O)₂—,        —NR₁′—CR₁′R₁′—C(O)—NR₁′—, —CR₁′R₁′—C(O)—NR₁′—,        —NR₁′—C(O)—CR₁′═CR₁′—CR₁′R₁′—, —NR₁′—C(═N—CN)—NR₁′—,        —NR₁′—C(O)—CR₁′R₁′—O—, —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—O—,        —NR₁′—S(O)₂—CR₁′R₁′—, —NR₁′—S(O)₂—CR₁′R₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—; —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—,        —NR₁′—C(S)—NR₁′—CR₁′R₁′—CR₁′R₁′—, —NR₁′—C(O)—O—,

-   -   R³¹ is selected from an optionally substituted monocyclic or        bicyclic aryl, or an with the proviso that when R¹⁹ is

-   -    Z₁₀, Z₁₁, Z₁₂ and Z₁₃ are each CH, and R²¹ is —NHC(O)—, R³¹ is        not an optionally substituted phenyl.

In certain embodiments, the compounds of Structural Formula (XI) havethe following values:

-   -   each of X₇, X₈, X₉ and X₁₀ is independently selected from N,        CR²⁰, or CR₁′, wherein:        -   each R²⁰ is independently selected from H or a solubilizing            group;        -   each R₁′ is independently selected from H or optionally            substituted C₁-C₃ straight or branched alkyl;        -   one of X₇, X₈, X₉ and X₁₀ is N and the others are selected            from CR²⁰ or CR₁′; and        -   zero to one R²⁰ is a solubilizing group;    -   R¹⁹ is selected from:

-   -   wherein:        -   each Z₁₀, Z₁₁, Z₁₂ and Z₁₃ is independently selected from N,            CR²⁰, or CR₁′; and        -   each Z₁₄, Z₁₅ and Z₁₆ is independently selected from N,            NR₁′, S, O, CR²⁰, or CR₁′, wherein:        -   zero to two of Z₁₀, Z₁₁, Z₁₂ or Z₁₃ are N;        -   at least one of Z₁₄, Z₁₅ and Z₁₆ is N, NR₁′, S or O;        -   zero to one of Z₁₄, Z₁₅ and Z₁₆ is S or O;        -   zero to two of Z₁₄, Z₁₅ and Z₁₆ are N or NR₁′;        -   zero to one R²⁰ is a solubilizing group;        -   zero to one R₁′ is an optionally substituted C₁-C₃ straight            or branched alkyl; and    -   R²¹ is selected from —NR₁′—C(O)—, —NR₁′—S(O)₂—,        —NR₁′—C(O)—NR₁′—, —NR₁′—C(S)—NR₁′—, —NR₁′—C(S)—NR₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—NR₁′—, —NR₁′—C(═NR₁′)—NR₁′—, —C(O)—NR₁′—,        —C(O)—NR₁′—S(O)₂—, —NR₁′—, —CR₁′R₁′—, —NR₁′—C(O)—CR₁′═CR₁′—,        —NR₁′—S(O)₂—NR₁′—, —NR₁′—C(O)—NR₁′—S(O)₂—,        —NR₁′—CR₁′R₁′—C(O)—NR₁′—, —CR₁′R₁′—C(O)—NR₁′—,        —NR₁′—C(O)—CR₁′═CR₁′—CR₁′R₁′—, —NR₁′—C(═N—CN)—NR₁′—,        —NR₁′—C(O)—CR₁′R₁′—O—, —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—O—,        —NR₁′—S(O)₂—CR₁′R₁′—, —NR₁′—S(O)₂—CR₁′R₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—, —NR₁′—C(S)—NR₁′—CR₁′R₁′—CR₁′R₁′—,        —NR₁′—C(O)—O— or —NR₁′—C(O)—CR₁′R₁′—; and    -   R³¹ is selected from an optionally substituted monocyclic or        bicyclic aryl, or an optionally substituted monocyclic or        bicyclic heteroaryl, with the proviso that:    -   when X₇ is N, R¹⁹ is

-   -    and each of Z₁₀, Z₁₁, Z₁₂ and Z₁₃ is independently selected        from CR²⁰, or CR₁′, then:        -   a) at least one of X₈, X₉ or X₁₀ is C—(C₁-C₃ straight or            branched alkyl) or C-(solubilizing group); or        -   b) at least one of Z₁₀, Z₁₁, Z₁₂ and Z₁₃ is CR²⁰, wherein            R²⁰ is a solubilizing group.

In a further aspect, the invention provides compounds of StructuralFormula (XIII):

-   or a salt thereof, wherein:    -   R₁′ is selected from H or optionally substituted C₁-C₃ straight        or branched alkyl;    -   R²¹ is selected from —NR₁′—C(O)—, —NR₁′—S(O)₂—,        —NR₁′—C(O)—NR₁′—, —NR₁′—C(S)—NR₁′—, —NR₁′—C(S)—NR₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—NR₁′—, —NR₁′—C(═NR₁′)—NR₁′—, —C(O)—NR₁′—,        —C(O)—NR₁′—S(O)₂—, —NR₁′—, —CR₁′R₁′—, —NR₁′—C(O)—CR₁′═CR₁′—,        —NR₁′—S(O)₂—NR₁′—, —NR₁′—C(O)—NR₁′—S(O)₂—,        —NR₁′—CR₁′R₁′—C(O)—NR₁′—, —CR₁′R₁′—C(O)—NR₁′—,        —NR₁′—C(O)—CR₁′═CR₁′—CR₁′R₁′—, —NR₁′—C(═N—CN)—NR₁′—,        —NR₁′—C(O)—CR₁′R₁′—O—, —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—O—,        —NR₁′—S(O)₂—CR₁′R₁′—, —NR₁′—S(O)₂—CR₁′R₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—; —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—,        —NR₁′—C(S)—NR₁′—CR₁′R₁′—CR₁′R₁′—, —NR₁′—C(O)—O—,

-   -   R³¹ is selected from an optionally substituted monocyclic or        bicyclic aryl, or an optionally substituted monocyclic or        bicyclic heteroaryl, with the provisos that:    -   when R²¹ is —NH—C(O)—, R³¹ is not unsubstituted furyl,        5-bromofuryl, unsubstituted phenyl, phenyl monosubstituted with        halo or methyl, 3-or 4-methoxyphenyl, 4-butoxyphenyl,        4-t-butylphenyl, 3-trifluoromethylphenyl, 2-benzoylphenyl, 2-or        4-ethoxyphenyl, 2,3-, 2,4-, 3,4-, or 3,5-dimethoxyphenyl,        3,4,5-trimethoxyphenyl, 2,4-or 2-6 difluorophenyl,        3,4-dioxymethylene phenyl, 3,4-or 3,5-dimethylphenyl,        2-chloro-5-bromophenyl, 2-methoxy-5-chlorophenyl, unsubstituted        quinolinyl, thiazolyl substituted simultaneously with methyl and        phenyl, or ethoxy-substituted pyridinyl;    -   when R²¹ is —NH—C(O)—CH(CH₂—CH₃)—, R³¹ is not unsubstituted        phenyl;    -   when R²¹ is —NH—C(O)—CH₂—, R³¹ is not unsubstituted phenyl,        3-methylphenyl, 4-chlorophenyl, 4-ethoxyphenyl, 4-fluorophenyl        or 4-methoxyphenyl;    -   when R²¹ is —NH—C(O)—CH₂—O—, R³¹ is not unsubstituted phenyl or        4-chlorophenyl; and    -   when R²¹ is —NH—S(O)₂—, R³¹ is not 3,4-dioxymethylene phenyl,        2,4,5-trimethylphenyl, 2,4,6-trimethylphenyl, 2,4-or        3,4-dimethylphenyl, 2,5-difluorophenyl, 2,5-or        3,4-dimethoxyphenyl, fluorophenyl, 4-chlorophenyl,        4-bromophenyl, 4-ethylphenyl, 4-methylphenyl,        3-methyl-4-methoxyphenyl, unsubstituted phenyl, unsubstituted        pyridinyl, unsubstituted thienyl, chloro-substituted thienyl, or        methyl-substituted benzothiazolyl.

In one aspect, the invention provides sirtuin-modulating compounds ofStructural Formula (XIV):

-   or a salt thereof, wherein:    -   each of R²³ and R²⁴ is independently selected from H, —CH₃ or a        solubilizing group;    -   R²⁵ is selected from H, or a solubilizing group; and    -   R¹⁹ is selected from:

-   -   wherein:        -   each Z₁₀, Z₁₁, Z₁₂ and Z₁₃ is independently selected from N,            CR²⁰, or CR₁′; and        -   each Z₁₄, Z₁₅ and Z₁₆ is independently selected from N,            NR₁′, S, O, CR²⁰, or CR₁′, wherein:        -   zero to two of Z₁₀, Z₁₁, Z₁₂ or Z₁₃ are N;        -   at least one of Z₁₄, Z₁₅ and Z₁₆ is N, NR₁′, O or S;        -   zero to one of Z₁₄, Z₁₅ and Z₁₆ is S or O;        -   zero to two of Z₁₄, Z₁₅ and Z₁₆ are N or NR₁′;        -   zero to one R²⁰ is a solubilizing group; and        -   zero to one R₁′ is an optionally substituted C₁-C₃ straight            or branched alkyl;        -   each R²⁰ is independently selected from H or a solubilizing            group;    -   R²¹ is selected from —NR₁′—C(O)—, —NR₁′—S(O)₂—,        —NR₁′—C(O)—NR₁′—, —NR₁′—C(S)—NR₁′—, —NR₁′—C(S)—NR₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—NR₁′—, —NR₁′—C(═NR₁′)—NR₁′—, —C(O)—NR₁′—,        —C(O)—NR₁′—S(O)₂—, —NR₁′—, —CR₁′R₁′—, —NR₁′—C(O)—CR₁′═CR₁′—,        —NR₁′—S(O)₂—NR₁′—, —NR₁′—C(O)—NR₁′—S(O)₂—,        —NR₁′—CR₁′R₁′—C(O)—NR₁′—, —CR₁′R₁′—C(O)—NR₁′—,        —NR₁′—C(O)—CR₁′═CR₁′—CR₁′R₁′—, —NR₁′—C(═N—CN)—NR₁′—,        —NR₁′—C(O)—CR₁′R₁′—O—, —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—O—,        —NR₁′—S(O)₂—CR₁′R₁′—, —NR₁′—S(O)₂—CR₁′R₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—; —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—,        —NR₁′—C(S)—NR₁′—CR₁′R₁′—CR₁′R₁′—, —NR₁′—C(O)—O—,

-   -   each R₁′ is independently selected from H or optionally        substituted C₁-C₃ straight or branched alkyl; and    -   R³¹ is selected from an optionally substituted monocyclic or        bicyclic aryl, or an optionally substituted monocyclic or        bicyclic heteroaryl,        -   wherein when R¹⁹ is

-   -   -    R²¹ is —NH—C(O)— and R²⁵ is —H, R³¹ is not an optionally            substituted phenyl group, and wherein said compound is not            2-chloro-N-[3-[3-(cyclohexylamino)imidazo[1,2-a]pyridin-2-yl]phenyl]-4-nitrobenzamide.

In another aspect, the invention provides sirtuin-modulating compoundsof Structural Formula (XV):

-   or a salt thereof, wherein:    -   R²¹ is selected from —NR₁′—C(O)—, —NR₁′—S(O)₂—,        —NR₁′—C(O)—NR₁′—, —NR₁′—C(S)—NR₁′—, —NR₁′—C(S)—NR₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—NR₁′—, —NR₁′—C(═NR₁′)—NR₁′—, —C(O)—NR₁′—,        —C(O)—NR₁′—S(O)₂—, —NR₁′—, —CR₁′R₁′—, —NR₁′—C(O)—CR₁′═CR₁′—,        —NR₁′—S(O)₂—NR₁′—, —NR₁′—C(O)—NR₁′—S(O)₂—,        —NR₁′—CR₁′R₁′—C(O)—NR₁′—, —CR₁′R₁′—C(O)—NR₁′—,        —NR₁′—C(O)—CR₁′═CR₁′—CR₁′R₁′—, —NR₁′—C(═N—CN)—NR₁′—,        —NR₁′—C(O)—CR₁′R₁′—O—, —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—O—,        —NR₁′—S(O)₂—CR₁′R₁′—, —NR₁′—S(O)₂—CR₁′R₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—; —NR₁′—C(O)—CR₁′R₁′R₁′—CR₁′R₁′—,        —NR₁′—C(S)—NR₁′—CR₁′R₁′—CR₁′R₁′—, —NR₁′—C(O)—O—,

-   -   each R₁′ is independently selected from H or optionally        substituted C₁-C₃ straight or branched alkyl; and    -   R³² is selected from an optionally substituted bicyclic aryl, or        an optionally substituted monocyclic or bicyclic heteroaryl,        wherein:    -   when R²¹ is —NH—C(O)—, R³² is not unsubstituted 2-furyl,        2-(3-bromofuryl), unsubstituted 2-thienyl, unsubstituted        3-pyridyl, unsubstituted 4-pyridyl,

-   -   when R²¹ is —NR₁′—S(O)₂—, R³² is not unsubstituted 2-thienyl or        unsubstituted naphthyl.

In yet another aspect, the invention provides sirtuin-modulatingcompounds of Structural Formula (XVI):

-   or a salt thereof, wherein:    -   R²¹ is selected from —NR₁′—C(O)—, —NR₁′—S(O)₂—,        —NR₁′—C(O)—NR₁′—, —NR₁′—C(S)—NR₁′—, —NR₁′—C(S)—NR₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—NR₁′—, —NR₁′—C(═NR₁′)—NR₁′—, —C(O)—NR₁′—,        —C(O)—NR₁′—S(O)₂—, —NR₁′—, —CR₁′R₁′—, —NR₁′—C(O)—CR₁′═CR₁′—,        —NR₁′—S(O)₂—NR₁′—, —NR₁′—C(O)—NR₁′—S(O)₂—,        —NR₁′—CR₁′R₁′—C(O)—NR₁′—, —CR₁′R₁′—C(O)—NR₁′—,        —NR₁′—C(O)—CR₁′═CR₁′—CR₁′R₁′—, —NR₁′—C(═N—CN)—NR₁′—,        —NR₁′—C(O)—CR₁′R₁′—O—, —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—O—,        —NR₁′—S(O)₂—CR₁′R₁′—, —NR₁′—S(O)₂—CR₁′R₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—; —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—,        —NR₁′—C(S)—NR₁′—CR₁′R₁′—CR₁′R₁′—, —NR₁′—C(O)—O—,

-   -   each R₁′ is independently selected from H or optionally        substituted C₁-C₃ straight or branched alkyl; and    -   R³³ is an optionally substituted phenyl, wherein:    -   when R²¹ is —NH—C(O)—, R³³ is a substituted phenyl other than        phenyl singly substituted with halo, methyl, nitro or methoxy;        2-carboxyphenyl; 4-n-pentylphenyl; 4-ethoxyphenyl;        2-carboxy-3-nitrophenyl; 2-chloro-4-nitrophenyl;        2-methoxy-5-ethylphenyl; 2,4-dimethoxyphenyl;        3,4,5-trimethoxyphenyl; 2,4 dichlorophenyl; 2,6-difluorophenyl;        3,5-dinitrophenyl; or 3,4-dimethylphenyl;    -   when R²¹ is —NR₁′—C(O)—CR₁′R₁′— or —NH—C(O)—CH(CH₃)—O, R³³ is a        substituted phenyl;    -   when R²¹ is —NH—C(O)—CH₂, R³³ is not unsubstituted phenyl,        4-methoxyphenyl; 3,4-dimethoxyphenyl or 4-chlorophenyl;    -   when R²¹ is —NH—C(O)—CH₂—O, R³³ is not        2,4-bis(1,1-dimethylpropyl)phenyl;    -   when R²¹ is —NH—C(O)—NH—, R³³ is not 4-methoxyphenyl; and    -   when R²¹ is —NH—S(O)₂—, R³³ is a substituted phenyl other than        3-methylphenyl, 3-trifluoromethylphenyl, 2,4,5-or        2,4,6-trimethylphenyl, 2,4-or 3,4-dimethylphenyl, 2,5-or        3,4-dimethoxyphenyl, 2,5-dimethoxy-4-chlorophenyl,        3,6-dimethoxy, 4-methylphenyl, 2,5-or 3,4-dichlorophenyl,        2,5-diethoxyphenyl, 2-methyl -5-nitrophenyl,        2-ethoxy-5-bromophenyl, 2-methoxy-5-bromophenyl,        2-methoxy-3,4-dichlorophenyl, 2-methoxy-4-methyl-5-bromophenyl,        3,5-dinitro-4-methylphenyl, 3-methyl-4-methoxyphenyl,        3-nitro-4-methylphenyl, 3-methoxy-4-halophenyl,        3-methoxy-5-chlorophenyl, 4-n-butoxyphenyl, 4-halophenyl,        4-ethylphenyl, 4-methylphenyl, 4-nitrophenyl, 4-ethoxyphenyl,        4-acetylaminophenyl, 4-methoxyphenyl, 4-t -butylphenyl, or        para-biphenyl.

In a further ascept, the invention provides sirtuin-modulating compoundsof Structural Formula (XVII):

-   or a salt thereof, wherein:    -   each of R²³ and R²⁴ is independently selected from H or —CH₃,        wherein at least one of R²³ and R²⁴ is H; and    -   R²⁹ is phenyl substituted with:    -   a) two —O—CH₃ groups;    -   b) three —O—CH₃ groups located at the 2, 3 and 4 positions; or    -   c) one —N(CH₃)₂ group; and;    -   d) when R²³ is CH₃, one —O—CH₃ group at the 2 or 3 position,        wherein R²⁹ is optionally additionally substituted with a        solubilizing group.

In one aspect, the invention provides sirtuin-modulating compounds ofStructural Formula (XVIII):

-   or a salt thereof, wherein    -   R¹⁹ is selected from:

-   -   wherein:        -   each Z₁₀, Z₁₁, Z₁₂ and Z₁₃ is independently selected from N,            CR²⁰, or CR₁′; and        -   each Z₁₄, Z₁₅ and Z₁₆ is independently selected from N,            NR₁′, S, O, CR²⁰, or CR₁′,    -   wherein:    -   zero to two of Z₁₀, Z₁₁, Z₁₂ or Z₁₃ are N;    -   at least one of Z₁₄, Z₁₅ and Z₁₆ is N, NR₁′, S or O;    -   zero to one of Z₁₄, Z₁₅ and Z₁₆ is S or O;    -   zero to two of Z₁₄, Z₁₅ and Z₁₆ are N or NR₁′;    -   zero to one R²⁰ is a solubilizing group; and    -   zero to one R₁′ is an optionally substituted C₁-C₃ straight or        branched alkyl;    -   each R²⁰ is independently selected from H or a solubilizing        group;    -   R²¹ is selected from —NR₁′—C(O)—, —NR₁′—S(O)₂—,        —NR₁′—C(O)—NR₁′—, —NR₁′—C(S)—NR₁′—, —NR₁′—C(S)—NR₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—NR₁′—, —NR₁′—C(═NR₁′)—NR₁′—, —C(O)—NR₁′—,        —C(O)—NR₁′—S(O)₂—, —NR₁′—, —CR₁′R₁′—, —NR₁′—C(O)—CR₁′═CR₁′—,        —NR₁′—S(O)₂—NR₁′—, —NR₁′—C(O)—NR₁′—S(O)₂—,        —NR₁′—CR₁′R₁′—C(O)—NR₁′—, —CR₁′R₁′—C(O)—NR₁′—,        —NR₁′—C(O)—CR₁′═CR₁′—CR₁′R₁′—, —NR₁′—C(═N—CN)—NR₁′—,        —NR₁′—C(O)—CR₁′R₁′—O—, —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—O—,        —NR₁′—S(O)₂—CR₁′R₁′—, —NR₁′—S(O)₂—CR₁′R₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—; —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—,        —NR₁′—C(S)—NR₁′—CR₁′R₁′—CR₁′R₁′—, —NR₁′—C(O)—O—,

-   -    wherein each R₁′ is independently selected from H or optionally        substituted C₁-C₃ straight or branched alkyl; and        -   R³¹ is selected from an optionally substituted monocyclic or            bicyclic aryl, or an optionally substituted monocyclic or            bicyclic heteroaryl, with the proviso that when R¹⁹ is

-   -   -    Z₁₀, Z₁₁, Z₁₂ and Z₁₃ are each CH, R¹⁰ is H, and R²¹ is            —NHC(O)—, R³¹ is not an optionally substituted phenyl.

In another aspect, the invention provides sirtuin-modulating compoundsof Structural Formula (XX):

-   or a salt thereof, wherein    -   R¹⁹ is selected from:

-   -   wherein:        -   each Z₁₀, Z₁₁, Z₁₂ and Z₁₃ is independently selected from N,            CR²⁰, or CR₁′; and        -   each Z₁₄, Z₁₅ and Z₁₆ is independently selected from N,            NR₁′, S, O, CR²⁰, or CR₁′,    -   wherein:        -   zero to two of Z₁₀, Z₁₁, Z₁₂ or Z₁₃ are N;        -   at least one of Z₁₄, Z₁₅ and Z₁₆ is N, NR₁′, O or S;        -   zero to one of Z₁₄, Z₁₅ and Z₁₆ is S or O;        -   zero to two of Z₁₄, Z₁₅ and Z₁₆ are N or NR₁′;        -   zero to one R²⁰ is a solubilizing group; and        -   zero to one R₁′ is an optionally substituted C₁-C₃ straight            or branched alkyl;    -   each R²⁰ is independently selected from H or a solubilizing        group;    -   R^(20a) is independently selected from H or a solubilizing        group;    -   R²¹ is selected from —NR₁′—C(O)—, —NR₁′—S(O)₂—,        —NR₁′—C(O)—NR₁′—, —NR₁′—C(S)—NR₁′—, —NR₁′—C(S)—NR₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—NR₁′—, —NR₁′—C(═NR₁′)—NR₁′—, —C(O)—NR₁′—,        —C(O)—NR₁′—S(O)₂—, —NR₁′—, —CR₁′R₁′—, —NR₁′—C(O)—CR₁′═CR₁′—,        —NR₁′—S(O)₂—NR₁′—, —NR₁′—C(O)—NR₁′—S(O)₂—,        —NR₁′—CR₁′R₁′—C(O)—NR₁′—, —CR₁′R₁′—C(O)—NR₁′—,        —NR₁′—C(O)—CR₁′═CR₁′—CR₁′R₁′—, —NR₁′—C(═N—CN)—NR₁′—,        —NR₁′—C(O)—CR₁′R₁′—O—, —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—O—,        —NR₁′—S(O)₂—CR₁′R₁′—, —NR₁′—S(O)₂—CR₁′R₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—; —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—,        —NR₁′—C(S)—NR₁′—CR₁′R₁′—CR₁′R₁′—, —NR₁′—C(O)—O—,

-   -   wherein        -   each R₁′ is independently selected from H or optionally            substituted C₁-C₃ straight or branched alkyl; and    -   R³¹ is selected from an optionally substituted monocyclic or        bicyclic aryl, or an optionally substituted monocyclic or        bicyclic heteroaryl, wherein when R¹⁹ is

-   -    and Z₁₀, Z₁₁, Z₁₂ and Z₁₃ are each CH, R^(20a) is a        solubilizing group.

In yet another aspect, the invention provides sirtuin-modulatingcompounds of Structural Formula (XXI):

-   or a salt thereof, wherein    -   R²¹ is selected from —NR₁′—C(O)—, —NR₁′—S(O)₂—,        —NR₁′—C(O)—NR₁′—, —NR₁′—C(S)—NR₁′—, —NR₁′—C(S)—NR₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—NR₁′—, —NR₁′—C(═NR₁′)—NR₁′—, —C(O)—NR₁′—,        —C(O)—NR₁′—S(O)₂—, —NR₁′—, —CR₁′R₁′—, —NR₁′—C(O)—CR₁′═CR₁′—,        —NR₁′—S(O)₂—NR₁′—, —NR₁′—C(O)—NR₁′—S(O)₂—,        —NR₁′—CR₁′R₁′—C(O)—NR₁′—, —CR₁′R₁′—C(O)—NR₁′—,        —NR₁′—C(O)—CR₁′═CR₁′—CR₁′R₁′—, —NR₁′—C(═N—CN)—NR₁′—,        —NR₁′—C(O)—CR₁′R₁′—O—, —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—O—,        —NR₁′—S(O)₂—CR₁′R₁′—, —NR₁′—S(O)₂—CR₁′R₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—; —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—,        —NR₁′—C(S)—NR₁′—CR₁′R₁′—CR₁′R₁′—, —NR₁′—C(O)—O—,

-   -   wherein        -   each R₁′ is independently selected from H or optionally            substituted C₁-C₃ straight or branched alkyl; and    -   R³² is an optionally substituted monocyclic or bicyclic        heteroaryl, or an optionally substituted bicyclic aryl, wherein:    -   when R²¹ is —NH—C(O)—CH₂—, R³² is not unsubstituted thien-2-yl;    -   when R²¹ is —NH—C(O)—, R³² is not furan-2-yl, 5-bromofuran        -2-yl, or 2-phenyl-4-methylthiazol-5-yl;    -   when R²¹ is —NH—S(O)₂—, R³² is not unsubstituted naphthyl or        5-chlorothien-2-yl.

In a further aspect, the invention provides sirtuin-modulating compoundsof Structural Formula (XXII):

-   or a salt thereof, wherein:    -   R²¹ is selected from —NR₁′—C(O)—, —NR₁′—S(O)₂—,        —NR₁′—C(O)—NR₁′—, —NR₁′—C(S)—NR₁′—, —NR₁′—C(S)—NR₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—NR₁′—, —NR₁′—C(═NR₁′)—NR₁′—, —C(O)—NR₁′—,        —C(O)—NR₁′—S(O)₂—, —NR₁′—, —CR₁′R₁′—, —NR₁′—C(O)—CR₁′═CR₁′—,        —NR₁′—S(O)₂—NR₁′—, —NR₁′—C(O)—NR₁′—S(O)₂—,        —NR₁′—CR₁′R₁′—C(O)—NR₁′—, —CR₁′R₁′—C(O)—NR₁′—,        —NR₁′—C(O)—CR₁′═CR₁′—CR₁′R₁′—, —NR₁′—C(═N—CN)—NR₁′—,        —NR₁′—C(O)—CR₁′R₁′—O—, —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—O—,        —NR₁′—S(O)₂—CR₁′R₁′—, —NR₁′—S(O)₂—CR₁′R₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—; —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—,        —NR₁′—C(S)—NR₁′—CR₁′R₁′—CR₁′R₁′—, —NR₁′—C(O)—O—,

-   -    wherein each R₁′ is independently selected from H or optionally        substituted C₁-C₃ straight or branched alkyl; and    -   R³³ is an optionally substituted phenyl, wherein:    -   when R²¹ is —NR₁′—C(O)—, R₁′ is not H;    -   when R²¹ is —NH—C(O)—CH₂ or —NH—C(O)—CH₂—O—, R³³ is not        unsubstituted phenyl or 4-halophenyl; and    -   when R²¹ is —NH—S(O)₂—, R³³ is not unsubstituted phenyl, 2,4-or        3,4-dimethylphenyl, 2,4-dimethyl-5-methoxyphenyl,        2-methoxy-3,4-dichlorophenyl, 2-methoxy,        5-bromophenyl-3,4-dioxyethylenephenyl, 3,4-dimethoxyphenyl,        3,4-dichlorophenyl, 3,4-dimethylphenyl, 3-or 4-methylphenyl,        4-alkoxyphenyl, 4-phenoxyphenyl, 4-halophenyl, 4-biphenyl, or        4-acetylaminophenyl.

In one aspect, the invention provides sirtuin-modulating compounds ofStructural Formula (XXII):

-   or a salt thereof wherein:    -   R²¹ is selected from —NH—C(O)—, or —NH—C(O)—CH₂—; and    -   R³³ is phenyl substituted with    -   a) one —N(CH₃)₂ group;    -   b) one CN group at the 3 position;    -   c) one —S(CH₃) group; or    -   d)

-   -    bridging the 3 and 4 positions.

In another aspect, the invention provides sirtuin-modulating compoundsof Structural Formula (XXIII):

-   or a salt thereof, wherein:    -   each R²⁰ and R^(20a) is independently selected from H or a        solubilizing group;    -   each R₁′, R₁″ and R₁′″ is independently selected from H or        optionally substituted C₁-C₃ straight or branched alkyl;    -   R²¹ is selected from —NR₁′—C(O)—, —NR₁′—S(O)₂—,        —NR₁′—C(O)—NR₁′—, —NR₁′—C(S)—NR₁′—, —NR₁′—C(S)—NR₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—NR₁′—, —NR₁′—C(═NR₁′)—NR₁′—,        —NR₁′—C(O)—CR₁′═CR₁′—, —NR₁′—S(O)₂—NR₁′—,        —NR₁′—C(O)—NR₁′—S(O)₂—, —NR₁′—CR₁′R₁′—C(O)—NR₁′—,        —NR₁′—C(O)—CR₁′═CR₁′—CR₁′R₁′—, —NR₁′—C(═N—CN)—NR₁′—,        —NR₁′—C(O)—CR₁′R₁′—O—, —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—O—,        —NR₁′—S(O)₂—CR₁′R₁′—, —NR₁′—S(O)₂—CR₁′R₁′—CR₁′R₁′—, or        —NR₁′—C(O)—CR₁′R₁′—; and    -   R³¹ is selected from an optionally substituted monocyclic or        bicyclic aryl, or an optionally substituted monocyclic or        bicyclic heteroaryl, with the provisos that:    -   when R²¹ is —NH—C(O)—, R³¹ is not is not 3,5-dinitrophenyl,        4-butoxyphenyl,

-   -   when R²¹ is —NH—C(O)— and each of R²⁰, R^(20a), R₁′, R₁″ and        R₁′″ is hydrogen, R³¹ is not

-   -    unsubstituted phenyl, 2-or 4-nitrophenyl, 2,4-dinitrophenyl,        2-or 4-chlorophenyl, 2-bromophenyl, 4-fluorophenyl,        2,4-dichlorophenyl, 2-carboxyphenyl, 2-azidophenyl, 2-or        4-aminophenyl, 2-acetamidophenyl, 4-methylphenyl, or        4-methoxyphenyl;    -   when R²¹ is —NH—C(O)—, R₁″ is methyl; and each of R²⁰, R^(20a),        R₁′ and R₁′″ is hydrogen, R³¹ is not 2-methylaminophenyl,

-   -   when R²¹ is —NH—C(O)—CH₂— or NH—C(S)—NH—, and each of R²⁰,        R^(20a), R₁′, R₁″ and R₁′″ is hydrogen, R³¹ is not unsubstituted        phenyl;    -   when R²¹ is —NH—S(O)₂—, R₁″ is hydrogen or methyl, and each of        R²⁰, R^(20a), R₁′ and R₁′″ is hydrogen, R³¹ is not        4-methylphenyl; and    -   when R²¹ is —NH—S(O)₂—, R^(20a) is hydrogen or —CH₂—N(CH₂CH₃)₂,        and each of R²⁰, R₁′, R₁″ and R₁′″ is hydrogen, R³¹ is not

In a particular aspect, the invention provides sirtuin-modulatingcompounds of Structural Formula (XXIII):

-   or a salt thereof, wherein:    -   each R²⁰ and R^(20a) is independently selected from H or a        solubilizing group;    -   each R₁′, R₁″ and R₁′″ is independently selected from H or        optionally substituted C₁-C₃ straight or branched alkyl;    -   R²¹ is selected from —NR₁′—C(O)—, —NR₁′—S(O)₂—,        —NR₁′—C(O)—NR₁′—, —NR₁′—C(S)—NR₁′—, —NR₁′—C(S)—NR₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—NR₁′, —NR₁′—C(═NR₁′)—NR₁′—,        —NR₁′—C(O)—CR₁′═CR₁′—, —NR₁′—S(O)₂—NR₁′—,        —NR₁′—C(O)—NR₁′—S(O)₂—, —NR₁′—CR₁′R₁′—C(O)—NR₁′—,        —NR₁′—C(O)—CR₁′═CR₁′—CR₁′R₁′—, —NR₁′—C(═N—CN)—NR₁′—,        —NR₁′—C(O)—CR₁′R₁′—O—, —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—O—,        —NR₁′—S(O)₂—CR₁′R₁′—, —NR₁′—S(O)₂—CR₁′R₁′—CR₁′R₁′—, or        —NR₁′—C(O)—CR₁′R₁′—; and    -   R³¹ is selected from an optionally substituted monocyclic or        bicyclic aryl, or an optionally substituted monocyclic or        bicyclic heteroaryl,    -   wherein:        -   i) at least one R²⁰ is a solubilizing group or at least one            R₁′″ is an optionally substituted C₁-C₃ straight or branched            alkyl or both; or        -   ii) R^(20a) is a solubilizing group other than            CH₂—N(CH₂CH₃)₂.

In yet another aspect, the invention provides sirtuin-modulatingcompounds of Structural Formula (XXIV):

-   or a salt thereof, wherein:    -   each R²⁰ and R^(20a) is independently selected from H or a        solubilizing group;    -   each R₁′, R₁″ and R₁′″ is independently selected from H or        optionally substituted C₁-C₃ straight or branched alkyl;    -   R²¹ is selected from —NR²³—C(O)—, —NR₁′—S(O)₂—,        —NR₁′—C(O)—NR₁′—, —NR₁′—C(S)—NR₁′—, —NR₁′—C(S)—NR₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—NR₁′—, —NR₁′—C(═NR₁′)—NR₁′—,        —NR₁′—C(O)—CR₁′═CR₁′—, —NR₁′—S(O)₂—NR₁′—,        —NR₁′—C(O)—NR₁′—S(O)₂—, —NR₁′—CR₁′R₁′—C(O)—NR₁′—,        —NR₁′—C(O)—CR₁′═CR₁′—CR₁′R₁′—, —NR₁′—C(═N—CN)—NR₁′—,        —NR₁′—C(O)—CR₁′R₁′—O—, —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—O—,        —NR₁′—S(O)₂—CR₁′R₁′—, —NR₁′—S(O)₂—CR₁′R₁′—CR₁′R₁′—, or        —NR₁′—C(O)—CR₁′R₁′—; and    -   R³¹ is selected from an optionally substituted monocyclic or        bicyclic aryl, or an optionally substituted monocyclic or        bicyclic heteroaryl, with the provisos that:    -   when R²¹ is —NH—C(O)—CH₂—, R³¹ is not 2-methylphenyl, or        3,4-dimethoxyphenyl;    -   when R²¹ is —NH—C(O)—CH═CH—, R³¹ is not 2-chlorophenyl;    -   when R²¹ is —NH—C(O)—NH—, R³¹ is not unsubstituted        benzimidazolyl;    -   when R²¹ is —NH—S(O)₂—, and each of R²⁰, R^(20a), R₁′, R₁″ and        R₁′″ is hydrogen, R³¹ is not unsubstituted phenyl,        4-chlorophenyl, 4-methylphenyl, or 4-acetoamidophenyl;    -   when R²¹ is —NH—S(O)₂—, each of R₁′ and R₁′″ is methyl or        hydrogen, and each of R²⁰, R^(20a), and R₁″ is hydrogen, R³¹ is        not 4-nitrophenyl;    -   when R²¹ is —NH—C(O)—CH₂—O—, R₁′″ is methyl or hydrogen, and        each of R²⁰, R^(20a), R₁′, and R₁″ is hydrogen, R³¹ is not 2,3-,        2,5-, 2,6-, 3,4-or 3,5-dimethylphenyl, 2,4-dichloromethyl,        2,4-dimethyl-6-bromophenyl, 2-or 4-chlorophenyl,        2-(1-methylpropyl)phenyl, 5-methyl-2-(1-methylethyl)phenyl, 2-or        4-methylphenyl, 2,4-dichloro-6-methylphenyl, nitrophenyl,        2,4-dimethyl-6-nitrophenyl, 2-or 4-methoxyphenyl,        4-acetyl-2-methoxyphenyl, 4-chloro-3,5-dimethylphenyl,        3-ethylphenyl, 4-bromophenyl, 4-cyclohexyphenyl,        4-(1-methylpropyl)phenyl, 4-(1-methylethyl)phenyl,        4-(1,1-dimethylethyl)phenyl, or unsubstituted phenyl;    -   when R²¹ is —NH—C(O)—CH₂—, R₁′″ is methyl or hydrogen, and each        of R²⁰, R^(20a), R₁′, and R₁″ is hydrogen, R³¹ is not        unsubstituted naphthyl, 4-chlorophenyl, 4-nitrophenyl,        4-methoxyphenyl, unsubstituted phenyl, unsubstituted thienyl

-   -   when R²¹ is —NH—C(O)—CH₂—, R₁′ is methyl, and each of R²⁰,        R^(20a), R₁″, and R₁′″ is hydrogen, R³¹ is not unsubstituted        phenyl;    -   when R²¹ is —NH—C(O)—CH═CH, R₁′″ is methyl or hydrogen, and each        of R²⁰, R^(20a), R₁′, and R₁″ is hydrogen, R³¹ is not        unsubstituted furyl, nitrophenyl-substituted furyl,        2,4-dichlorophenyl, 3,5-dichloro-2-methoxyphenyl, 3-or        4-nitrophenyl, 4-methoxyphenyl, unsubstituted phenyl, or        nitro-substituted thienyl;    -   when R²¹ is —NH—C(O)—CH(CH₂CH₃)—, and each of R²⁰, R^(20a), R₁′,        R₁″, and R₁′″ is hydrogen, R³¹ is not unsubstituted phenyl;    -   when R²¹ is —NH—C(O)—CH(CH₃)—O—, R₁′″ is methyl or hydrogen, and        each of R₂₀, R^(20a), R₁′, and R₁″ is hydrogen, R³¹ is not        2,4-dichlorophenyl.

In a particular aspect, the invention provides sirtuin-modulatingcompounds of Structural Formula (XXIV):

-   or a salt thereof, wherein:    -   each R²⁰ and R^(20a) is independently selected from H or a        solubilizing group and at least one of R²⁰ and R^(20a) is a        solubilizing group;    -   each R₁′, R₁″ and R₁′″ is independently selected from H or        optionally substituted C₁-C₃ straight or branched alkyl;    -   R²¹ is selected from —NR²³—C(O)—, —NR₁′—S(O)₂—,        —NR₁′—C(O)—NR₁′—, —NR₁′—C(S)—NR₁′—, —NR₁′—C(S)—NR₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—NR₁′—, —NR₁′—C(═NR₁′)—NR₁′—,        —NR₁′—C(O)—CR₁′═CR₁′—, —NR₁′—S(O)₂—NR₁′—,        —NR₁′—C(O)—NR₁′—S(O)₂—, —NR₁′—CR₁′R₁′—C(O)—NR₁′—,        —NR₁′—C(O)—CR₁′═CR₁′—CR₁′R₁′—, —NR₁′—C(═N—CN)—NR₁′—,        —NR₁′—C(O)—CR₁′R₁′—O—, —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—O—,        —NR₁′—S(O)₂—CR₁′R₁′—, —NR₁₁′—S(O)₂—CR₁′R₁′—CR₁′R₁′—, or        —NR₁′—C(O)—CR₁′R₁′—, wherein R²³ is an optionally substituted        C₁-C₃ straight or branched alkyl; and    -   R³¹ is selected from an optionally substituted monocyclic or        bicyclic aryl, or an optionally substituted monocyclic or        bicyclic heteroaryl.

In a further aspect, the invention provides sirtuin-modulating compoundsof Structural Formula (XXV):

-   or a salt thereof, wherein:    -   each R²⁰ and R^(20a) is independently selected from H or a        solubilizing group, wherein at least one of R²⁰ and R^(20a) is a        solubilizing group;    -   each R₁′, R₁″ and R₁′″ is independently selected from H or        optionally substituted C₁-C₃ straight or branched alkyl; and    -   R³² is an optionally substituted phenyl.

In one aspect, the invention provides sirtuin-modulating compounds ofStructural Formula (XXVI):

-   or a salt thereof, wherein:    -   each R²⁰ and R^(20a) is independently selected from H or a        solubilizing group;    -   each R₁′, R₁″ and R₁′″ is independently selected from H or        optionally substituted C₁-C₃ straight or branched alkyl; and    -   R³³ is selected from an optionally substituted heteroaryl or an        optionally substituted bicyclic aryl, with the provisos that:    -   when each of R₁′ and R₁′″ is hydrogen or methyl and each of R¹″,        R₂₀ and R_(20a) is hydrogen, R³³ is not        5,6,7,8-tetrahydronaphthyl, unsubstituted benzofuryl,        unsubstituted benzothiazolyl, chloro- or nitro-substituted        benzothienyl, unsubstituted furyl, phenyl-, bromo- or        nitro-substituted furyl, dimethyl-substituted isoxazolyl,        unsubstituted naphthyl, 5-bromonaphthyl, 4-methylnaphthyl, 1- or        3-methoxynaphthyl, azo-substituted naphthyl, unsubstituted        pyrazinyl, S-methyl-substituted pyridyl, unsubstituted pyridyl,        thienyl- or phenyl-substituted quinolinyl, chloro-, bromo- or        nitro-substituted thienyl, unsubstituted thienyl, or

In a particular aspect, the invention provides sirtuin-modulatingcompounds of Structural Formula (XXVI):

-   or a salt thereof, wherein:    -   each R²⁰ and R^(20a) is independently selected from H or a        solubilizing group, wherein at least one of R₂₀ or R^(20a) is a        solubilizing group;    -   each R₁′, R₁″ and R₁′″ is independently selected from H or        optionally substituted C₁-C₃ straight or branched alkyl; and    -   R³³ is selected from an optionally substituted heteroaryl or an        optionally substituted bicyclic aryl.

In another aspect, the invention provides sirtuin-modulating compoundsof Structural Formula (XXVII):

-   or a salt thereof, wherein:    -   each R²⁰ and R^(20a) is independently selected from H or a        solubilizing group;    -   each R₁′ and R₁″ is independently selected from H or optionally        substituted C₁-C₃ straight or branched alkyl;    -   R¹⁹ is selected from:

-   -   wherein:        -   each Z₁₀, Z₁₁, Z₁₂ and Z₁₃ is independently selected from N,            CR²⁰, or CR₁′; and        -   each Z₁₄, Z₁₅ and Z₁₆ is independently selected from N,            NR₁′, S, O, CR²⁰, or CR₁′, wherein:        -   zero to two of Z₁₀, Z₁₁, Z₁₂ or Z₁₃ are N;        -   at least one of Z₁₄, Z₁₅ and Z₁₆ is N, NR₁′, S or O;        -   zero to one of Z₁₄, Z₁₅ and Z₁₆ is S or O;        -   zero to two of Z₁₄, Z₁₅ and Z₁₆ are N or NR₁′;        -   zero to one R²⁰ is a solubilizing group;        -   zero to one R₁′ is an optionally substituted C₁-C₃ straight            or branched alkyl; and        -   R²¹ is selected from —NR₁′—C(O)—, —NR₁′—S(O)₂—,            —NR₁′—C(O)—NR₁′—, —NR₁′—C(S)—NR₁′—,            —NR₁′—C(S)—NR₁′—CR₁′R₁′—, —NR₁′—C(O)—CR₁′R₁′—NR₁′—,            —NR₁′—C(═NR₁′)—NR₁′—, —C(O)—NR₁′—, —C(O)—NR₁′—S(O)₂—,            —NR₁′—, —CR₁′R₁′—, —NR₁′—C(O)—CR₁′═CR₁′—, —NR₁′—S(O)₂—NR₁′—,            —NR₁′—C(O)—NR₁′—S(O)₂—, —NR₁′—CR₁′R₁′—C(O)—NR₁′—,            —CR₁′R₁′—C(O)—NR₁′—, —NR₁′—C(O)—CR₁′═CR₁′—CR₁′R₁′—,            —NR₁′—C(═N—CN)—NR₁′—, —NR₁′—C(O)—CR₁′R₁′—O—,            —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—O—, —NR₁′—S(O)₂—CR₁′R₁′—,            —NR₁′—S(O)₂—CR₁′R₁′—CR₁′R₁′—, or —NR₁′—C(O)—CR₁′R₁′—; and    -   R³¹ is selected from an optionally substituted monocyclic or        bicyclic aryl, or an optionally substituted monocyclic or        bicyclic heteroaryl,        -   provided that when R²¹ is —NH—C(O)— and R¹⁹ is

-   -   -    R³¹ is not unsubstituted pyridyl, 2,6-dimethoxyphenyl,            3,4,5-trimethoxyphenyl or unsubstituted furyl.

In a particular aspect, the invention provides sirtuin-modulatingcompounds of Structural Formula (XXVII):

-   or a salt thereof, wherein:    -   each R²⁰ and R^(20a) is independently selected from H or a        solubilizing group;    -   each R₁′ and R₁″ is independently selected from H or optionally        substituted C₁-C₃ straight or branched alkyl;

R¹⁹ is selected from:

-   -   wherein:    -   each Z₁₀, Z₁₁, Z₁₂ and Z₁₃ is independently selected from N,        CR²⁰, or CR₁′; and    -   each Z₁₄, Z₁₅ and Z₁₆ is independently selected from N, NR₁′, S,        O, CR²⁰, or CR₁′, wherein:    -   zero to two of Z₁₀, Z₁₁, Z₁₂ or Z₁₃ are N;    -   at least one of Z₁₄, Z₁₅ and Z₁₆ is N, NR₁′, S or O;    -   zero to one of Z₁₄, Z₁₅ and Z₁₆ is S or O;    -   zero to two of Z₁₄, Z₁₅ and Z₁₆ are N or NR₁′;    -   zero to one R²⁰ is a solubilizing group;    -   zero to one R₁′ is an optionally substituted C₁-C₃ straight or        branched alkyl; and    -   R²¹ is selected from —NR₁′—C(O)—, —NR₁′—S(O)₂—,        —NR₁′—C(O)—NR₁′—, —NR₁′—C(S)—NR₁′—, —NR₁′—C(S)—NR₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—NR₁′—, —NR₁′—C(═NR₁′)—NR₁′—, —C(O)—NR₁′—,        —C(O)—NR₁′—S(O)₂—, —NR₁′—, —CR₁′R₁′—, —NR₁′—C(O)—CR₁′═CR₁′—,        —NR₁′—S(O)₂—NR₁′—, —NR₁′—C(O)—NR₁′—S(O)₂—,        —NR₁′—CR₁′R₁′—C(O)—NR₁′—, —CR₁′R₁′—C(O)—NR₁′—,        —NR₁′—C(O)—CR₁′═CR₁′—CR₁′R₁′—, —NR₁′—C(═N—CN)—NR₁′—,        —NR₁′—C(O)—CR₁′R₁′—O—, —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—O—,        —NR₁′—S(O)₂—CR₁′R₁′—, —NR₁′—S(O)₂—CR₁′R₁′—CR₁′R₁′—, or        —NR₁′—C(O)—CR₁′R₁′—; and    -   R³¹ is selected from an optionally substituted monocyclic or        bicyclic aryl, or an optionally substituted monocyclic or        bicyclic heteroaryl, with the provisos that:    -   when R²¹ is —NH—C(O)—, R¹⁹ is not pyrazolyl;    -   when R²¹ is —NH—, and R¹⁹ is thiazolyl, R³¹ is not optionally        substituted phenyl or optionally substituted pyridyl;    -   when R²¹ is —NH—C(O)—CH₂—, and R¹⁹ is pyrazolyl, R³¹ is not        unsubstituted indolyl or unsubstituted phenyl; ‘when R²¹ is        —NH—C(O)—CH₂—, and R¹⁹ is

-   -    R³¹ is not 2-methylphenyl or 3,4-dimethoxyphenyl;    -   when R²¹ is —NH—C(O)—CH═CH—, and R¹⁹ is

-   -    R³¹ is not 2-chlorophenyl;    -   when R²¹ is —NH—C(O)—NH—, and R¹⁹ is pyrazolyl, R³¹ is not        unsubstituted isoxazolyl, unsubstituted naphthyl, unsubstituted        phenyl, 2,6-difluorophenyl, 2,5-dimethylphenyl,        3,4-dichlorophenyl, or 4-chlorophenyl;    -   when R²¹ is —NH—C(O)—NH—, and R¹⁹ is

-   -    R³¹ is not unsubstituted benzimidazolyl;    -   when R²¹ is —NH—, and R¹⁹ is pyrazolyl, R³¹ is not unsubstituted        pyridyl;    -   when R^(20a) is a solubilizing group, R¹⁹ is 1-methylpyrrolyl        and R²¹ is —NH—C(O)—, R³¹ is not unsubstituted phenyl,        unsubstituted furyl, unsubstituted pyrrolyl, unsubstituted        pyrazolyl, unsubstituted isoquinolinyl, unsubstituted        benzothienyl, chloro-substituted benzothienyl,        2-fluoro-4-chlorophenyl or phenyl singly substituted with a        solubilizing group;    -   when R^(20a) is a solubilizing group, R¹⁹ is thienyl and R²¹ is        —NH—C(O)—, R³¹ is not unsubstituted phenyl;    -   when R^(20a) is a solubilizing group, R¹⁹ is methylimidazolyl        and R²¹ is —NH—C(O)—, R³¹ is not        1-methyl-4-(1,1-dimethylethyloxycarbonylamino)pyrrol-2-yl or        phenyl singly substituted with a solubilizing group;    -   when R²¹ is —NH— and R¹⁹ is pyridyl, oxadiazolyl or        thiadiazolyl, R³¹ is not unsubstituted phenyl, 3-methoxyphenyl        or 4-methoxyphenyl;    -   when R²¹ is —NH—C(O)— and R¹⁹ is thiazolyl or pyrimidinyl, R³¹        is not unsubstituted phenyl;    -   when R²¹ is —NH—C(O)— and R¹⁹ is

-   -    R³¹ is not unsubstituted pyridyl, unsubstituted thienyl,        unsubstituted phenyl, 2-methylphenyl, 4-fluorophenyl,        4-methoxyphenyl, 4-methylphenyl, 3,4-dioxyethylenephenyl,        3-acetylamino-4-methylphenyl,        3-[(6-amino-1-oxohexyl)amino]-4-methylphenyl,        3-amino-4-methylphenyl, 2,6-dimethoxyphenyl,        3,5-dimethoxyphenyl, 3-halo -4-methoxyphenyl,        3-nitro-4-methylphenyl, 4-propoxyphenyl, 3,4,5-trimethoxyphenyl        or unsubstituted furyl;    -   when R²¹ is —NH—C(O)— and R¹⁹ is

-   -    R³¹ is not 3,5-dinitrophenyl, 4-butoxyphenyl,

In a more particular embodiment, the invention providessirtuin-modulating compounds of Structural Formula (XXVII):

-   or a salt thereof, wherein:    -   each R²⁰ and R^(20a) is independently selected from H or a        solubilizing group;    -   each R₁′ and R₁″ is independently selected from H or optionally        substituted C₁-C₃ straight or branched alkyl;    -   R¹⁹ is selected from:

-   -   wherein:        -   each Z₁₀, Z₁₁, Z₁₂ and Z₁₃ is independently selected from N,            CR²⁰, or CR₁′; and    -   each Z₁₄, Z₁₅ and Z₁₆ is independently selected from N, NR₁′, S,        O, CR²⁰, or CR₁′, wherein:    -   one to two of Z₁₀, Z₁₁, Z₁₂ or Z₁₃ are N;    -   at least one of Z₁₄, Z₁₅ and Z₁₆ is N, NR₁′, S or O;    -   zero to one of Z₁₄, Z₁₅ and Z₁₆ is S or O;    -   zero to two of Z₁₄, Z₁₅ and Z₁₆ are N or NR₁′;    -   zero to one R²⁰ is a solubilizing group;    -   zero to one R₁′″ is an optionally substituted C₁-C₃ straight or        branched alkyl; and    -   R²¹ is selected from —NR₁′—C(O)—, —NR₁′—S(O)₂—,        —NR₁′—C(O)—NR₁′—, —NR₁′—C(S)—NR₁′—, —NR₁′—C(S)—NR₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—NR₁′—, —NR₁′—C(═NR₁′)—NR₁′—,        —NR₁′—C(O)—CR₁′═CR₁′—, —NR₁′—S(O)₂—NR₁′—,        —NR₁′—C(O)—NR₁′—S(O)₂—, —NR₁′—CR₁′R₁′—C(O)—NR₁′—,        —NR₁′—C(O)—CR₁′═CR₁′—CR₁′R₁′—, —NR₁′—C(═N—CN)—NR₁′—,        —NR₁′—C(O)—CR₁′R₁′—O—, —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—O—,        —NR₁′—S(O)₂—CR₁′R₁′—, —NR₁′—S(O)₂—CR₁′R₁′—CR₁′R₁′—, or        —NR₁′—C(O)—CR₁′R₁′—; and    -   R³¹ is selected from an optionally substituted monocyclic or        bicyclic aryl, or an optionally substituted monocyclic or        bicyclic heteroaryl, with the provisos that:    -   when R²¹ is —NH—C(O)—, R¹⁹ is not pyrazolyl;    -   when R²¹ is —NH—C(O)—CH₂—, and R¹⁹ is pyrazolyl, R³¹ is not        unsubstituted indolyl or unsubstituted phenyl;    -   when R²¹ is —NH—C(O)—NH—, and R¹⁹ is pyrazolyl, R³¹ is not        unsubstituted isoxazolyl, unsubstituted naphthyl, unsubstituted        phenyl, 2,6-difluorophenyl; 2,5-dimethylphenyl;        3,4-dichlorophenyl; or 4-chlorophenyl;    -   when R^(20a) is a solubilizing group, R¹⁹ is 1-methylpyrrolyl        and R²¹ is —NH—C(O)—, R³¹ is not unsubstituted phenyl;        unsubstituted furyl; unsubstituted pyrrolyl; unsubstituted        pyrazolyl; unsubstituted isoquinolinyl; unsubstituted        benzothienyl; chloro-substituted benzothienyl;        2-fluoro-4-chlorophenyl or phenyl singly substituted with a        solubilizing group;    -   when R^(20a) is a solubilizing group, R¹⁹ is thienyl and R²¹ is        —NH—C(O)—, R³¹ is not unsubstituted phenyl;    -   when R^(20a) is a solubilizing group, R¹⁹ is methylimidazolyl        and R²¹ is —NH—C(O)—, R³¹ is not        1-methyl-4-(1,1-dimethylethyloxycarbonylamino)pyrrol-2-yl or        phenyl singly substituted with a solubilizing group; and    -   when R²¹ is —NH—C(O)— and R¹⁹ is thiazolyl or pyrimidinyl, R³¹        is not unsubstituted phenyl.

In yet another aspect, the invention provides compounds of StructuralFormula (XXVIII):

-   or a salt thereof, wherein:    -   each R²⁰ and R^(20a) is independently selected from H or a        solubilizing group;    -   each R₁′ and R₁″ is independently selected from H or optionally        substituted C₁-C₃ straight or branched alkyl;    -   R²⁹ is selected from:

-   -    wherein:    -   each Z₁₀, Z₁₀, Z₁₂ and Z₁₃ is independently selected from N,        CR²⁰, or CR₁′, wherein one of Z₁₀, Z₁₁, Z₁₂ or Z₁₃ is N; and    -   zero to one R²⁰ is a solubilizing group;    -   zero to one R₁′″ is an optionally substituted C₁-C₃ straight or        branched alkyl; and    -   R²¹ is selected from —NR₁′—C(O)—, —NR₁′—S(O)₂—,        —NR₁′—C(O)—NR₁′—, —NR₁′—C(S)—NR₁′—, —NR₁′—C(S)—NR₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—NR₁′—, —NR₁′—C(═NR₁′)—NR₁′—,        —NR₁′—C(O)—CR₁′═CR₁′—, —NR₁′—S(O)₂—NR₁′—,        —NR₁′—C(O)—NR₁′—S(O)₂—, —NR₁′—CR₁′R₁′—C(O)—NR₁′—,        —NR₁′—C(O)—CR₁′═CR₁′—CR₁′R₁′—, —NR₁′—C(═N—CN)—NR₁′—,        —NR₁′—C(O)—CR₁′R₁′—O—, —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—O—,        —NR₁′—S(O)₂—CR₁′R₁′—, —NR₁′—S(O)₂—CR₁′R₁′—CR₁′R₁′—, or        —NR₁′—C(O)—CR₁′R₁′—; and    -   R³¹ is selected from an optionally substituted monocyclic or        bicyclic aryl, or an optionally substituted monocyclic or        bicyclic heteroaryl.

Also provided are pharmaceutical compositions comprising one or morecompounds of Formulas (I)-(XXVIII) or a salt, prodrug or metabolitethereof.

In another aspect, the invention provides methods for usingsirtuin-modulating compounds, or compostions comprisingsirtuin-modulating compounds. In certain embodiments, sirtuin-modulatingcompounds that increase the level and/or activity of a sirtuin proteinmay be used for a variety of therapeutic applications including, forexample, increasing the lifespan of a cell, and treating and/orpreventing a wide variety of diseases and disorders including, forexample, diseases or disorders related to aging or stress, diabetes,obesity, neurodegenerative diseases, chemotherapeutic inducedneuropathy, neuropathy associated with an ischemic event, oculardiseases and/or disorders, cardiovascular disease, blood clottingdisorders, inflammation, and/or flushing, etc. Sirtuin-modulatingcompounds that increase the level and/or activity of a sirtuin proteinmay also be used for treating a disease or disorder in a subject thatwould benefit from increased mitochondrial activity, for enhancingmuscle performance, for increasing muscle ATP levels, or for treating orpreventing muscle tissue damage associated with hypoxia or ischemia. Inother embodiments, sirtuin-modulating compounds that decrease the leveland/or activity of a sirtuin protein may be used for a variety oftherapeutic applications including, for example, increasing cellularsensitivity to stress, increasing apoptosis, treatment of cancer,stimulation of appetite, and/or stimulation of weight gain, etc. Asdescribed further below, the methods comprise administering to a subjectin need thereof a pharmaceutically effective amount of asirtuin-modulating compound.

In certain aspects, the sirtuin-modulating compounds may be administeredalone or in combination with other compounds, including othersirtuin-modulating compounds, or other therapeutic agents.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 shows a schematic of the Cellular ATP Assay described in Example5.

FIG. 2 shows a dose-response curve for ATP levels in cells followingresveratrol treatment.

DETAILED DESCRIPTION

1. Definitions

As used herein, the following terms and phrases shall have the meaningsset forth below. Unless defined otherwise, all technical and scientificterms used herein have the same meaning as commonly understood to one ofordinary skill in the art.

The singular forms “a,” “an,” and “the” include plural reference unlessthe context clearly dictates otherwise.

The term “agent” is used herein to denote a chemical compound, a mixtureof chemical compounds, a biological macromolecule (such as a nucleicacid, an antibody, a protein or portion thereof, e.g., a peptide), or anextract made from biological materials such as bacteria, plants, fungi,or animal (particularly mammalian) cells or tissues. The activity ofsuch agents may render it suitable as a “therapeutic agent” which is abiologically, physiologically, or pharmacologically active substance (orsubstances) that acts locally or systemically in a subject.

The term “bioavailable” when referring to a compound is art-recognizedand refers to a form of a compound that allows for it, or a portion ofthe amount of compound administered, to be absorbed by, incorporated to,or otherwise physiologically available to a subject or patient to whomit is administered.

“Biologically active portion of a sirtuin” refers to a portion of asirtuin protein having a biological activity, such as the ability todeacetylate. Biologically active portions of a sirtuin may comprise thecore domain of sirtuins. Biologically active portions of SIRT1 havingGenBank Accession No. NP_(—)036370 that encompass the NAD+ bindingdomain and the substrate binding domain, for example, may includewithout limitation, amino acids 62-293 of GenBank Accession No.NP_(—)036370, which are encoded by nucleotides 237 to 932 of GenBankAccession No. NM_(—)012238. Therefore, this region is sometimes referredto as the core domain. Other biologically active portions of SIRT1, alsosometimes referred to as core domains, include about amino acids 261 to447 of GenBank Accession No. NP_(—)036370, which are encoded bynucleotides 834 to 1394 of GenBank Accession No. NM_(—)012238; aboutamino acids 242 to 493 of GenBank Accession No. NP_(—)036370, which areencoded by nucleotides 777 to 1532 of GenBank Accession No.NM_(—)012238; or about amino acids 254 to 495 of GenBank Accession No.NP_(—)036370, which are encoded by nucleotides 813 to 1538 of GenBankAccession No. NM_(—)012238.

The term “companion animals” refers to cats and dogs. As used herein,the term “dog(s)” denotes any member of the species Canis familiaris, ofwhich there are a large number of different breeds. The term “cat(s)”refers to a feline animal including domestic cats and other members ofthe family Felidae, genus Felis.

The terms “comprise” and “comprising” are used in the inclusive, opensense, meaning that additional elements may be included.

The term “conserved residue” refers to an amino acid that is a member ofa group of amino acids having certain common properties. The term“conservative amino acid substitution” refers to the substitution(conceptually or otherwise) of an amino acid from one such group with adifferent amino acid from the same group. A functional way to definecommon properties between individual amino acids is to analyze thenormalized frequencies of amino acid changes between correspondingproteins of homologous organisms (Schulz, G. E. and R. H. Schirmer.,Principles of Protein Structure, Springer-Verlag). According to suchanalyses, groups of amino acids may be defined where amino acids withina group exchange preferentially with each other, and therefore resembleeach other most in their impact on the overall protein structure(Schulz, G. E. and R. H. Schirmer, Principles of Protein Structure,Springer-Verlag). One example of a set of amino acid groups defined inthis manner include: (i) a charged group, consisting of Glu and Asp,Lys, Arg and His, (ii) a positively-charged group, consisting of Lys,Arg and His, (iii) a negatively-charged group, consisting of Glu andAsp, (iv) an aromatic group, consisting of Phe, Tyr and Trp, (v) anitrogen ring group, consisting of His and Trp, (vi) a large aliphaticnonpolar group, consisting of Val, Leu and Ile, (vii) a slightly-polargroup, consisting of Met and Cys, (viii) a small-residue group,consisting of Ser, Thr, Asp, Asn, Gly, Ala, Glu, Gln and Pro, (ix) analiphatic group consisting of Val, Leu, Ile, Met and Cys, and (x) asmall hydroxyl group consisting of Ser and Thr.

“Diabetes” refers to high blood sugar or ketoacidosis, as well aschronic, general metabolic abnormalities arising from a prolonged highblood sugar status or a decrease in glucose tolerance. “Diabetes”encompasses both the type I and type II (Non Insulin Dependent DiabetesMellitus or NIDDM) forms of the disease. The risk factors for diabetesinclude the following factors: waistline of more than 40 inches for menor 35 inches for women, blood pressure of 130/85 mm/Hg or higher,triglycerides above 150 mg/dl, fasting blood glucose greater than 100mg/dl or high-density lipoprotein of less than 40 mg/dl in men or 50mg/dl in women.

A “direct activator” of a sirtuin is a molecule that activates a sirtuinby binding to it. A “direct inhibitor” of a sirtuin is a moleculeinhibits a sirtuin by binding to it.

The term “ED₅₀” is art-recognized. In certain embodiments, ED₅₀ meansthe dose of a drug which produces 50% of its maximum response or effect,or alternatively, the dose which produces a pre-determined response in50% of test subjects or preparations. The term “LD₅₀” is art-recognized.In certain embodiments, LD₅₀ means the dose of a drug which is lethal in50% of test subjects. The term “therapeutic index” is an art-recognizedterm which refers to the therapeutic index of a drug, defined asLD₅₀/ED₅₀.

The term “hyperinsulinemia” refers to a state in an individual in whichthe level of insulin in the blood is higher than normal.

The term “including” is used to mean “including but not limited to”.“Including” and “including but not limited to” are used interchangeably.

The term “insulin resistance” refers to a state in which a normal amountof insulin produces a subnormal biologic response relative to thebiological response in a subject that does not have insulin resistance.

An “insulin resistance disorder,” as discussed herein, refers to anydisease or condition that is caused by or contributed to by insulinresistance. Examples include: diabetes, obesity, metabolic syndrome,insulin-resistance syndromes, syndrome X, insulin resistance, high bloodpressure, hypertension, high blood cholesterol, dyslipidemia,hyperlipidemia, dyslipidemia, atherosclerotic disease including stroke,coronary artery disease or myocardial infarction, hyperglycemia,hyperinsulinemia and/or hyperproinsulinemia, impaired glucose tolerance,delayed insulin release, diabetic complications, including coronaryheart disease, angina pectoris, congestive heart failure, stroke,cognitive functions in dementia, retinopathy, peripheral neuropathy,nephropathy, glomerulonephritis, glomerulosclerosis, nephrotic syndrome,hypertensive nephrosclerosis some types of cancer (such as endometrial,breast, prostate, and colon), complications of pregnancy, poor femalereproductive health (such as menstrual irregularities, infertility,irregular ovulation, polycystic ovarian syndrome (PCOS)), lipodystrophy,cholesterol related disorders, such as gallstones, cholescystitis andcholelithiasis, gout, obstructive sleep apnea and respiratory problems,osteoarthritis, and prevention and treatment of bone loss, e.g.osteoporosis.

The term “livestock animals” refers to domesticated quadrupeds, whichincludes those being raised for meat and various byproducts, e.g., abovine animal including cattle and other members of the genus Bos, aporcine animal including domestic swine and other members of the genusSus, an ovine animal including sheep and other members of the genusOvis, domestic goats and other members of the genus Capra; domesticatedquadrupeds being raised for specialized tasks such as use as a beast ofburden, e.g., an equine animal including domestic horses and othermembers of the family Equidae, genus Equus.

The term “mammal” is known in the art, and exemplary mammals includehumans, primates, livestock animals (including bovines, porcines, etc.),companion animals (e.g., canines, felines, etc.) and rodents (e.g., miceand rats).

The term “naturally occurring form” when referring to a compound means acompound that is in a form, e.g., a composition, in which it can befound naturally. For example, since resveratrol can be found in redwine, it is present in red wine in a form that is naturally occurring. Acompound is not in a form that is naturally occurring if, e.g., thecompound has been purified and separated from at least some of the othermolecules that are found with the compound in nature. A “naturallyoccurring compound” refers to a compound that can be found in nature,i.e., a compound that has not been designed by man. A naturallyoccurring compound may have been made by man or by nature.

A “naturally occurring compound” refers to a compound that can be foundin nature, i.e., a compound that has not been designed by man. Anaturally occurring compound may have been made by man or by nature. Forexample, resveratrol is a naturally-occurring compound. A “non-naturallyoccurring compound” is a compound that is not known to exist in natureor that does not occur in nature.

“Obese” individuals or individuals suffering from obesity are generallyindividuals having a body mass index (BMI) of at least 25 or greater.Obesity may or may not be associated with insulin resistance.

The terms “parenteral administration” and “administered parenterally”are art -recognized and refer to modes of administration other thanenteral and topical administration, usually by injection, and includes,without limitation, intravenous, intramuscular, intraarterial,intrathecal, intracapsular, intraorbital, intracardiac, intradermal,intraperitoneal, transtracheal, subcutaneous, subcuticular,intra-articulare, subcapsular, subarachnoid, intraspinal, andintrasternal injection and infusion.

A “patient”, “subject”, “individual” or “host” refers to either a humanor a non-human animal.

The term “percent identical” refers to sequence identity between twoamino acid sequences or between two nucleotide sequences. Identity caneach be determined by comparing a position in each sequence which may bealigned for purposes of comparison. When an equivalent position in thecompared sequences is occupied by the same base or amino acid, then themolecules are identical at that position; when the equivalent siteoccupied by the same or a similar amino acid residue (e.g., similar insteric and/or electronic nature), then the molecules can be referred toas homologous (similar) at that position. Expression as a percentage ofhomology, similarity, or identity refers to a function of the number ofidentical or similar amino acids at positions shared by the comparedsequences. Expression as a percentage of homology, similarity, oridentity refers to a function of the number of identical or similaramino acids at positions shared by the compared sequences. Variousalignment algorithms and/or programs may be used, including FASTA,BLAST, or ENTREZ. FASTA and BLAST are available as a part of the GCGsequence analysis package (University of Wisconsin, Madison, Wis.), andcan be used with, e.g., default settings. ENTREZ is available throughthe National Center for Biotechnology Information, National Library ofMedicine, National Institutes of Health, Bethesda, Md. In oneembodiment, the percent identity of two sequences can be determined bythe GCG program with a gap weight of 1, e.g., each amino acid gap isweighted as if it were a single amino acid or nucleotide mismatchbetween the two sequences.

Other techniques for alignment are described in Methods in Enzymology,vol. 266: Computer Methods for Macromolecular Sequence Analysis (1996),ed. Doolittle, Academic Press, Inc., a division of Harcourt Brace & Co.,San Diego, Calif., USA. Preferably, an alignment program that permitsgaps in the sequence is utilized to align the sequences. TheSmith-Waterman is one type of algorithm that permits gaps in sequencealignments. See Meth. Mol. Biol. 70: 173-187 (1997). Also, the GAPprogram using the Needleman and Wunsch alignment method can be utilizedto align sequences. An alternative search strategy uses MPSRCH software,which runs on a MASPAR computer. MPSRCH uses a Smith-Waterman algorithmto score sequences on a massively parallel computer. This approachimproves ability to pick up distantly related matches, and is especiallytolerant of small gaps and nucleotide sequence errors. Nucleicacid-encoded amino acid sequences can be used to search both protein andDNA databases.

The term “pharmaceutically acceptable carrier” is art-recognized andrefers to a pharmaceutically-acceptable material, composition orvehicle, such as a liquid or solid filler, diluent, excipient, solventor encapsulating material, involved in carrying or transporting anysubject composition or component thereof. Each carrier must be“acceptable” in the sense of being compatible with the subjectcomposition and its components and not injurious to the patient. Someexamples of materials which may serve as pharmaceutically acceptablecarriers include: (1) sugars, such as lactose, glucose and sucrose; (2)starches, such as corn starch and potato starch; (3) cellulose, and itsderivatives, such as sodium carboxymethyl cellulose, ethyl cellulose andcellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7)talc; (8) excipients, such as cocoa butter and suppository waxes; (9)oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil,olive oil, corn oil and soybean oil; (10) glycols, such as propyleneglycol; (11) polyols, such as glycerin, sorbitol, mannitol andpolyethylene glycol; (12) esters, such as ethyl oleate and ethyllaurate; (13) agar; (14) buffering agents, such as magnesium hydroxideand aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17)isotonic saline; (18) Ringer's solution; (19) ethyl alcohol; (20)phosphate buffer solutions; and (21) other non-toxic compatiblesubstances employed in pharmaceutical formulations.

The terms “polynucleotide”, and “nucleic acid” are used interchangeably.They refer to a polymeric form of nucleotides of any length, eitherdeoxyribonucleotides or ribonucleotides, or analogs thereof.Polynucleotides may have any three-dimensional structure, and mayperform any function, known or unknown. The following are non-limitingexamples of polynucleotides: coding or non-coding regions of a gene orgene fragment, loci (locus) defined from linkage analysis, exons,introns, messenger RNA (mRNA), transfer RNA, ribosomal RNA, ribozymes,cDNA, recombinant polynucleotides, branched polynucleotides, plasmids,vectors, isolated DNA of any sequence, isolated RNA of any sequence,nucleic acid probes, and primers. A polynucleotide may comprise modifiednucleotides, such as methylated nucleotides and nucleotide analogs. Ifpresent, modifications to the nucleotide structure may be impartedbefore or after assembly of the polymer. The sequence of nucleotides maybe interrupted by non-nucleotide components. A polynucleotide may befurther modified, such as by conjugation with a labeling component. Theterm “recombinant” polynucleotide means a polynucleotide of genomic,cDNA, semi synthetic, or synthetic origin which either does not occur innature or is linked to another polynucleotide in a nonnaturalarrangement.

The term “prophylactic” or “therapeutic” treatment is art-recognized andrefers to administration of a drug to a host. If it is administeredprior to clinical manifestation of the unwanted condition (e.g., diseaseor other unwanted state of the host animal) then the treatment isprophylactic, i.e., it protects the host against developing the unwantedcondition, whereas if administered after manifestation of the unwantedcondition, the treatment is therapeutic (i.e., it is intended todiminish, ameliorate or maintain the existing unwanted condition or sideeffects therefrom).

The term “protecting group” is art-recognized and refers to temporarysubstituents that protect a potentially reactive functional group fromundesired chemical transformations. Examples of such protecting groupsinclude esters of carboxylic acids, silyl ethers of alcohols, andacetals and ketals of aldehydes and ketones, respectively. The field ofprotecting group chemistry has been reviewed by Greene and Wuts inProtective Groups in Organic Synthesis (2^(nd) ed., Wiley: New York,1991).

The term “pyrogen-free”, with reference to a composition, refers to acomposition that does not contain a pyrogen in an amount that would leadto an adverse effect (e.g., irritation, fever, inflammation, diarrhea,respiratory distress, endotoxic shock, etc.) in a subject to which thecomposition has been administered. For example, the term is meant toencompass compositions that are free of, or substantially free of, anendotoxin such as, for example, a lipopolysaccharide (LPS).

“Replicative lifespan” of a cell refers to the number of daughter cellsproduced by an individual “mother cell.” “Chronological aging” or“chronological lifespan,” on the other hand, refers to the length oftime a population of non-dividing cells remains viable when deprived ofnutrients. “Increasing the lifespan of a cell” or “extending thelifespan of a cell,” as applied to cells or organisms, refers toincreasing the number of daughter cells produced by one cell; increasingthe ability of cells or organisms to cope with stresses and combatdamage, e.g., to DNA, proteins; and/or increasing the ability of cellsor organisms to survive and exist in a living state for longer under aparticular condition, e.g., stress (for example, heatshock, osmoticstress, high energy radiation, chemically-induced stress, DNA damage,inadequate salt level, inadequate nitrogen level, or inadequate nutrientlevel). Lifespan can be increased by at least about 20%, 30%, 40%, 50%,60% or between 20% and 70%, 30% and 60%, 40% and 60% or more usingmethods described herein.

“Sirtuin-activating compound” refers to a compound that increases thelevel of a sirtuin protein and/or increases at least one activity of asirtuin protein. In an exemplary embodiment, a sirtuin-activatingcompound may increase at least one biological activity of a sirtuinprotein by at least about 10%, 25%, 50%, 75%, 100%, or more. Exemplarybiological activities of sirtuin proteins include deacetylation, e.g.,of histones and p53; extending lifespan; increasing genomic stability;silencing transcription; and controlling the segregation of oxidizedproteins between mother and daughter cells.

“Sirtuin-inhibiting compound” refers to a compound that decreases thelevel of a sirtuin protein and/or decreases at least one activity of asirtuin protein. In an exemplary embodiment, a sirtuin-inhibitingcompound may decrease at least one biological activity of a sirtuinprotein by at least about 10%, 25%, 50%, 75%, 100%, or more. Exemplarybiological activities of sirtuin proteins include deacetylation, e.g.,of histones and p53; extending lifespan; increasing genomic stability;silencing transcription; and controlling the segregation of oxidizedproteins between mother and daughter cells.

“Sirtuin-modulating compound” refers to a compound of Formulas(I)-(XXVI) as described herein. In exemplary embodiments, asirtuin-modulating compound may either up regulate (e.g., activate orstimulate), down regulate (e.g., inhibit or suppress) or otherwisechange a functional property or biological activity of a sirtuinprotein. Sirtuin-modulating compounds may act to modulate a sirtuinprotein either directly or indirectly. In certain embodiments, asirtuin-modulating compound may be a sirtuin-activating compound or asirtuin-inhibiting compound.

“Sirtuin protein” refers to a member of the sirtuin deacetylase proteinfamily, or preferably to the sir2 family, which include yeast Sir2(GenBank Accession No. P53685), C. elegans Sir-2.1 (GenBank AccessionNo. NP_(—)501912), and human SIRT1 (GenBank Accession No. NM_(—)012238and NP_(—)036370 (or AF083106)) and SIRT2 (GenBank Accession No.NM_(—)012237, NM_(—)030593, NP_(—)036369, NP_(—)085096, and AF083107)proteins. Other family members include the four additional yeastSir2-like genes termed “HST genes” (homologues of Sir two) HST1, HST2,HST3 and HST4, and the five other human homologues hSIRT3, hSIRT4,hSIRT5, hSIRT6 and hSIRT7, (Brachmann et al. (1995) Genes Dev. 9:2888and Frye et al. (1999) BBRC 260:273). Preferred sirtuins are those thatshare more similarities with SIRT1, i.e., hSIRT1, and/or Sir2 than withSIRT2, such as those members having at least part of the N -terminalsequence present in SIRT1 and absent in SIRT2 such as SIRT3 has.

“SIRT1 protein” refers to a member of the sir2 family of sirtuindeacetylases. In one embodiment, a SIRT1 protein includes yeast Sir2(GenBank Accession No. P53685), C. elegans Sir-2.1 (GenBank AccessionNo. NP_(—)501912), human SIRT1 (GenBank Accession No. NM_(—)012238 orNP_(—)036370 (or AF083106)), and human SIRT2 (GenBank Accession No.NM_(—)012237, NM_(—)030593, NP_(—)036369, NP_(—)085096, or AF083107)proteins, and equivalents and fragments thereof. In another embodiment,a SIRT1 protein includes a polypeptide comprising a sequence consistingof, or consisting essentially of, the amino acid sequence set forth inGenBank Accession Nos. NP_(—)036370, NP_(—)501912, NP_(—)085096,NP_(—)036369, or P53685. SIRT1 proteins include polypeptides comprisingall or a portion of the amino acid sequence set forth in GenBankAccession Nos. NP_(—)036370, NP_(—)501912, NP_(—)085096, NP_(—)036369,or P53685; the amino acid sequence set forth in GenBank Accession Nos.NP_(—)036370, NP_(—)501912, NP_(—)085096, NP_(—)036369, or P53685 with 1to about 2, 3, 5, 7, 10, 15, 20, 30, 50, 75 or more conservative aminoacid substitutions; an amino acid sequence that is at least 60%, 70%,80%, 90%, 95%, 96%, 97%, 98%, or 99% identical to GenBank Accession Nos.NP_(—)036370, NP_(—)501912, NP_(—)085096, NP_(—)036369, or P53685, andfunctional fragments thereof. Polypeptides of the invention also includehomologs (e.g., orthologs and paralogs), variants, or fragments, ofGenBank Accession Nos. NP_(—)036370, NP_(—)501912, NP_(—)085096,NP_(—)036369, or P53685.

“SIRT3 protein” refers to a member of the sirtuin deacetylase proteinfamily and/or to a homolog of a SIRT1 protein. In one embodiment, aSIRT3 protein includes human SIRT3 (GenBank Accession No. AAH01042,NP_(—)036371, or NP_(—)001017524) and mouse SIRT3 (GenBank Accession No.NP_(—)071878) proteins, and equivalents and fragments thereof. Inanother embodiment, a SIRT3 protein includes a polypeptide comprising asequence consisting of, or consisting essentially of, the amino acidsequence set forth in GenBank Accession Nos. AAH01042, NP_(—)036371,NP_(—)001017524, or NP_(—)071878. SIRT3 proteins include polypeptidescomprising all or a portion of the amino acid sequence set forth inGenBank Accession AAH01042, NP_(—)036371, NP_(—)001017524, orNP_(—)071878; the amino acid sequence set forth in GenBank AccessionNos. AAH01042, NP_(—)036371, NP_(—)001017524, or NP_(—)071878 with 1 toabout 2, 3, 5, 7, 10, 15, 20, 30, 50, 75 or more conservative amino acidsubstitutions; an amino acid sequence that is at least 60%, 70%, 80%,90%, 95%, 96%, 97%, 98%, or 99% identical to GenBank Accession Nos.AAH01042, NP_(—)036371, NP_(—)001017524, or NP_(—)071878, and functionalfragments thereof. Polypeptides of the invention also include homologs(e.g., orthologs and paralogs), variants, or fragments, of GenBankAccession Nos. AAH01042, NP_(—)036371, NP_(—)001017524, or NP_(—)071878.In one embodiment, a SIRT3 protein includes a fragment of SIRT3 proteinthat is produced by cleavage with a mitochondrial matrix processingpeptidase (MPP) and/or a mitochondrial intermediate peptidase (MIP).

The term “substantially homologous” when used in connection with aminoacid sequences, refers to sequences which are substantially identical toor similar in sequence with each other, giving rise to a homology ofconformation and thus to retention, to a useful degree, of one or morebiological (including immunological) activities. The term is notintended to imply a common evolution of the sequences.

The term “synthetic” is art-recognized and refers to production by invitro chemical or enzymatic synthesis.

The terms “systemic administration,” “administered systemically,”“peripheral administration” and “administered peripherally” areart-recognized and refer to the administration of a subject composition,therapeutic or other material other than directly into the centralnervous system, such that it enters the patient's system and, thus, issubject to metabolism and other like processes.

The term “therapeutic agent” is art-recognized and refers to anychemical moiety that is a biologically, physiologically, orpharmacologically active substance that acts locally or systemically ina subject. The term also means any substance intended for use in thediagnosis, cure, mitigation, treatment or prevention of disease or inthe enhancement of desirable physical or mental development and/orconditions in an animal or human.

The term “therapeutic effect” is art-recognized and refers to a local orsystemic effect in animals, particularly mammals, and more particularlyhumans caused by a pharmacologically active substance. The phrase“therapeutically-effective amount” means that amount of such a substancethat produces some desired local or systemic effect at a reasonablebenefit/risk ratio applicable to any treatment. The therapeuticallyeffective amount of such substance will vary depending upon the subjectand disease condition being treated, the weight and age of the subject,the severity of the disease condition, the manner of administration andthe like, which can readily be determined by one of ordinary skill inthe art. For example, certain compositions described herein may beadministered in a sufficient amount to produce a desired effect at areasonable benefit/risk ratio applicable to such treatment.

“Transcriptional regulatory sequence” is a generic term used throughoutthe specification to refer to DNA sequences, such as initiation signals,enhancers, and promoters, which induce or control transcription ofprotein coding sequences with which they are operable linked. Inpreferred embodiments, transcription of one of the recombinant genes isunder the control of a promoter sequence (or other transcriptionalregulatory sequence) which controls the expression of the recombinantgene in a cell-type which expression is intended. It will also beunderstood that the recombinant gene can be under the control oftranscriptional regulatory sequences which are the same or which aredifferent from those sequences which control transcription of thenaturally-occurring forms of genes as described herein.

“Treating” a condition or disease refers to curing as well asameliorating at least one symptom of the condition or disease.

A “vector” is a self-replicating nucleic acid molecule that transfers aninserted nucleic acid molecule into and/or between host cells. The termincludes vectors that function primarily for insertion of a nucleic acidmolecule into a cell, replication of vectors that function primarily forthe replication of nucleic acid, and expression vectors that functionfor transcription and/or translation of the DNA or RNA. Also includedare vectors that provide more than one of the above functions. As usedherein, “expression vectors” are defined as polynucleotides which, whenintroduced into an appropriate host cell, can be transcribed andtranslated into a polypeptide(s). An “expression system” usuallyconnotes a suitable host cell comprised of an expression vector that canfunction to yield a desired expression product.

The term “vision impairment” refers to diminished vision, which is oftenonly partially reversible or irreversible upon treatment (e.g.,surgery). Particularly severe vision impairment is termed “blindness” or“vision loss”, which refers to a complete loss of vision, vision worsethan 20/200 that cannot be improved with corrective lenses, or a visualfield of less than 20 degrees diameter (10 degrees radius).

2. Sirtuin Modulators

In one aspect, the invention provides novel sirtuin-modulating compoundsfor treating and/or preventing a wide variety of diseases and disordersincluding, for example, diseases or disorders related to aging orstress, diabetes, obesity, neurodegenerative diseases, ocular diseasesand disorders, cardiovascular disease, blood clotting disorders,inflammation, cancer, and/or flushing, etc. Sirtuin-modulating compoundsthat increase the level and/or activity of a sirtuin protein may also beused for treating a disease or disorder in a subject that would benefitfrom increased mitochondrial activity, for enhancing muscle performance,for increasing muscle ATP levels, or for treating or preventing muscletissue damage associated with hypoxia or ischemia. Other compoundsdisclosed herein may be suitable for use in a pharmaceutical compositionand/or one or more methods disclosed herein.

In one embodiment, sirtuin-modulating compounds of the invention arerepresented by Structural Formula (I):

-   or a salt thereof, where:    -   Ring A is optionally substituted; and    -   Ring B is substituted with at least one carboxy, substituted or        unsubstituted arylcarboxamine, substituted or unsubstituted        aralkylcarboxamine, substituted or unsubstituted heteroaryl        group, substituted or unsubstituted heterocyclylcarbonylethenyl,        or polycyclic aryl group or is fused to an aryl ring and is        optionally substituted by one or more additional groups.

In certain embodiments, Ring B is substituted with at least a carboxygroup.

In certain embodiments, Ring B is substituted with at least asubstituted or unsubstituted arylcarboxamine, a substituted orunsubstituted aralkylcarboxamine or a polycyclic aryl group.

In certain embodiments, Ring B is substituted with at least asubstituted or unsubstituted heteroaryl group or a substituted orunsubstituted heterocyclylcarbonylethenyl group.

In another embodiment, sirtuin-modulating compounds of the invention arerepresented by Structural Formula (II):

-   or a salt thereof, where:    -   Ring A is optionally substituted;    -   R₁, R₂, R₃ and R₄ are independently selected from the group        consisting of —H, halogen, —OR₅, —CN, —CO₂R₅, —OCOR₅, —OCO₂R₅,        —C(O)NR₅R₆, —OC(O)NR₅R₆, —C(O)R₅, —COR₅, —SR₅, —OSO₃H,        —S(O)_(n)R₅, —S(O)_(n)OR₅, —S(O)_(n)NR₅R₆, —NR₅R₆, —NR₅C(O)OR₆,        —NR₅C(O)R₆ and —NO₂;    -   R₅ and R₆ are independently —H, a substituted or unsubstituted        alkyl group, a substituted or unsubstituted aryl group or a        substituted or unsubstituted heterocyclic group; and

n is 1 or 2.

In a further embodiment, sirtuin-modulating compounds of the inventionare represented by Structural Formula (IIa):

-   or a salt thereof, where:    -   Ring A is optionally substituted;    -   R₁, R₂, R₃ and R₄ are independently selected from the group        consisting of —H, halogen, —OR₅, —CN, —CO₂R₅, —OCOR₅, —OCO₂R₅,        —C(O)NR₅R₆, —OC(O)NR₅R₆, —C(O)R₅, —COR₅, —SR₅, —OSO₃H,        —S(O)_(n)R₅, —S(O)_(n)OR₅, —S(O)_(n)NR₅R₆, —NR₅R₆, —NR₅C(O)OR₆,        —NR₅C(O)R₆ and —NO₂;    -   R₅ and R₆ are independently —H, a substituted or unsubstituted        alkyl group, a substituted or unsubstituted aryl group or a        substituted or unsubstituted heterocyclic group; and    -   n is 1 or 2.

In yet another embodiment, sirtuin-modulating compounds of the inventionare represented by Structural Formula (II):

-   or a salt thereof, where:    -   Ring A is optionally substituted;    -   R₁, R₂, R₃ and R₄ are independently selected from the group        consisting of —H, halogen, —OR₅, —CN, —CO₂R₅, —OCOR₅, —OCO₂R₅,        —C(O)NR₅R₆, —OC(O)NR₅R₆, —C(O)R₅, —COR₅, —SR₅, —OSO₃H,        —S(O)_(n)R₅, —S(O)_(n)OR₅, —S(O)_(n)NR₅R₆, —NR₅R₆, —NR₅C(O)OR₆,        —NR₅C(O)R₆ and —NO₂;    -   R₅ and R⁶ are independently —H, a substituted or unsubstituted        alkyl group, a substituted or unsubstituted aryl group or a        substituted or unsubstituted heterocyclic group; and    -   n is 1 or 2.

In certain embodiments, R₁, R₂, R₃ and R₄ in Structural Formulas(II)-(IIb) are independently selected from the group consisting of —H,—OR₅ and —SR₅, particularly —H and —OR₅ (e.g., —H, —OH, —OCH₃).

Ring A is preferably substituted. Suitable substituents include halogens(e.g., bromine), acyloxy groups (e.g., acetoxy), aminocarbonyl groups(e.g., arylaminocarbonyl such as substituted, particularlycarboxy-substituted, phenylaminocarbonyl groups) and alkoxy (e.g.,methoxy, ethoxy) groups.

In yet another aspect, the invention provides novel sirtuin-modulatingcompounds of Formula (If):

-   or a salt thereof, where:    -   Ring A is optionally substituted;    -   R₅ and R₆ are independently —H, a substituted or unsubstituted        alkyl group, a substituted or unsubstituted aryl group or a        substituted or unsubstituted heterocyclic group;    -   R₇, R₉, R₁₀ and R₁₁ are independently selected from the group        consisting of —H, halogen, —R₅, —OR₅, —CN, —CO₂R₅, —OCOR₅,        —OCO₂R₅, —C(O)NR₅R₆, —OC(O)NR₅R₆, —C(O)R₅, —COR₅, —SR₅, —OSO₃H,        —S(O)_(n)R₅, —S(O)_(n)OR₅, —S(O)_(n)NR₅R₆, —NR₅R₆, —NR₅C(O)OR₆,        —NR₅C(O)R₆ and —NO₂;    -   R₈ is a polycyclic aryl group; and    -   n is 1 or 2.

In certain embodiments, one or more of R₇, R₉, R₁₀ and R₁₁ are —H. Inparticular embodiments, R₇, R₉, R₁₀ and R₁₁ are each —H.

In certain embodiments, R₈ is a heteroaryl group, such as anoxazolo[4,5-b]pyridyl group. In particular embodiments, R₈ is aheteroaryl group and one or more of R₇, R₉, R₁₀ and R₁₁ are —H.

Ring A is preferably substituted. Suitable substituents include halogens(e.g., bromine), acyloxy groups (e.g., acetoxy), aminocarbonyl groups(e.g., arylaminocarbonyl, such as substituted, particularlycarboxy-substituted, phenylaminocarbonyl groups) and alkoxy (e.g.,methoxy, ethoxy) groups, particularly alkoxy groups. In certainembodiments, Ring A is substituted with at least one alkoxy or halogroup, particularly methoxy.

In certain embodiments, Ring A is optionally substituted with up to 3substituents independently selected from (C₁-C₃ straight or branchedalkyl), O—(C₁-C₃ straight or branched alkyl), N(C₁-C₃ straight orbranched alkyl)₂, halo, or a 5 to 6-membered heterocycle.

In certain embodiments, Ring A is not substituted with a nitrile orpyrrolidyl group.

In certain embodiments, R₈ is a substituted or unsubstituted bicyclicheteroaryl group, such as a bicyclic heteroaryl group that includes aring N atom and 1 to 2 additional ring heteroatoms independentlyselected from N, O or S. Preferably, R₈ is attached to the remainder ofthe compound by a carbon-carbon bond. In certain such embodiments, 2additional ring heteroatoms are present, and typically at least one ofsaid additional ring heteroatoms is O or S. In certain such embodiments,2 total ring nitrogen atoms are present (with zero or one O or Spresent), and the nitrogen atoms are typically each in a different ring.In certain such embodiments, R₈ is not substituted with acarbonyl-containing moiety, particularly when R₈ is thienopyrimidyl orthienopyridinyl.

In certain such embodiments, R₈ is selected from oxazolopyridyl,benzothienyl, benzofuryl, indolyl, quinoxalinyl, benzothiazolyl,benzoxazolyl, benzimidazolyl, quinolinyl, isoquinolinyl or isoindolyl.In certain such embodiments, R₈ is selected from thiazolopyridyl,imidazothiazolyl, benzoxazinonyl, or imidazopyridyl.

Particular examples of R₈, where

indicates attachment to the remainder of Structural Formula (III),include:

where up to 2 ring carbons not immediately adjacent to the indicatedattachment point are independently substituted with O—C₁-C₃ straight orbranched alkyl, C₁-C₃ straight or branched alkyl or halo, particularlyC₁-C₃ straight or branched alkyl or halo. In certain embodiments, R₈ is

In certain embodiments (e.g., when the modulator is a sirtuinactivator), R₈ is

and Ring A is optionally substituted with up to 3 substituentsindependently selected from (C₁-C₃ straight or branched alkyl), O—(C₁-C₃straight or branched alkyl), N(C₁-C₃ straight or branched alkyl)₂, halo,or a 5 to 6-membered heterocycle. In certain such embodiments, Ring A isnot simultaneously substituted at the 2-and 6-positions with O—(C₁-C₃straight or branched alkyl). In certain such embodiments, Ring A is notsimultaneously substituted at the 2-, 4-and 6-positions with O—(C₁-C₃straight or branched alkyl). In certain such embodiments, Ring A is notsimultaneously substituted at the 2-, 3-, and 4-positions with O—(C₁-C₃straight or branched alkyl). In certain such embodiments, Ring A is notsubstituted at the 4-position with a 5 to 6-membered heterocycle. Incertain such embodiments, Ring A is not singly substituted at the 3- or4-position (typically 4-position) with O—(C₁-C₃ straight or branchedalkyl). In certain such embodiments, Ring A is not substituted at the4-position with O—(C₁-C₃ straight or branched alkyl) and at the 2- or3-position with C₁-C₃ straight or branched alkyl.

In certain embodiments, R₈ is

and Ring A is optionally substituted with up to 3 substituentsindependently selected from (C₁-C₃ straight or branched alkyl), (C₁-C₃straight or branched haloalkyl, where a haloalkyl group is an alkylgroup substituted with one or more halogen atoms), O—(C₁-C₃ straight orbranched alkyl), N(C₁-C₃ straight or branched alkyl)₂, halo, or a 5 to6-membered heterocycle. In certain such embodiments, Ring A is notsingly substituted at the 3- or 4-position with O—(C₁-C₃ straight orbranched alkyl). In certain such embodiments, Ring A is not substitutedat the 4-position with O—(C₁-C₃ straight or branched alkyl) and at the2- or- or 3-position with C₁-C₃ straight or branched alkyl.

In certain embodiments, R₈ is

(e.g., where one or both halo is chlorine) and Ring A is optionallysubstituted with up to 3 substituents independently selected from (C₁-C₃straight or branched alkyl), O—(C₁-C₃ straight or branched alkyl),N(C₁-C₃ straight or branched alkyl)₂, halo, or a 5 to 6-memberedheterocycle, but not singly substituted at the 3-position with O—(C₁-C₃straight or branched alkyl).

In certain embodiments, such as when R has one of the values describedabove, Ring A is substituted with up to 3 substituents independentlyselected from chloro, methyl, O-methyl, N(CH₃)₂ or morpholino. Incertain such embodiments, R₈ is selected from

where up to 2 ring carbons not immediately adjacent to the indicatedattachment point are independently substituted with C₁-C₃ straight orbranched alkyl or halo; each of R₇, R₉, and R₁₁ is —H; and R₁₀ isselected from —H, —CH₂OH, —CO₂H, —CO₂CH₃, —CH₂-piperazinyl, CH₂N(CH₃)₂,—C(O)—NH—(CH₂)₂—N(CH₃)₂, or —C(O)-piperazinyl. In certain suchembodiments, when R₈ is

and Ring A is 3-dimethylaminophenyl, none of R₇, R₉, R₁₀ and R₁₁ is—CH₂—N(CH₃)₂ or —C(O)—NH—(CH₂)₂—N(CH₃)₂, and/or when R₈ is

and Ring A is 3,4 dimethoxyphenyl, none of R₇, R₉, R₁₀ and R₁₁ isC(O)OCH₃ or C(O)OH.

In certain embodiments, such as when R₈ has one of the values describedabove and/or Ring A is optionally substituted as described above, atleast one of R₇, R₉, R₁₀ and R₁₁ is —H. In certain such embodiments,each of R₇, R₉, R₁₀ and R₁₁ is —H.

In certain embodiments, R₇, R₉, R₁₀ or R₁₁ is selected from —C(O)OH,—N(CH₃)₂, —CH₂OH, —CH₂OCH₃, —CH₂-piperazinyl, —CH₂-methylpiperazinyl,—CH₂-pyrrolidyl, —CH₂-piperidyl, —CH₂-morpholino, —CH₂—N(CH₃)₂,—C(O)—NH—(CH₂)_(n)-piperazinyl, —C(O)—NH—(CH₂)_(n)-methylpiperazinyl,—C(O)—NH—(CH₂)_(n)-pyrrolidyl, —C(O)—NH—(CH₂)_(n)-morpholino,—C(O)—NH—(CH₂)_(n)-piperidyl, or —C(O)—NH—(CH₂)_(n)N(CH₃)₂, wherein n is1 or 2. In certain such embodiments, R₁₀ is selected from —C(O)OH,—N(CH₃)₂, —CH₂OH, —CH₂OCH₃, —CH₂-piperazinyl, —CH₂-methylpiperazinyl,—CH₂-pyrrolidyl, —CH₂-piperidyl, —CH₂-morpholino, —CH₂—N(CH₃)₂,—C(O)—NH—(CH₂)_(n)-piperazinyl, —C(O)—NH—(CH₂), methylpiperazinyl,—C(O)—NH—(CH₂)_(n)-pyrrolidyl, —C(O)—NH—(CH₂)_(n)-morpholino,—C(O)—NH—(CH₂)_(n)-piperidyl, or —C(O)—NH—(CH₂)_(n)—N(CH₃)₂, wherein nis 1 or 2, and each of R₇, R₉, and R₁₁ is H.

In certain embodiments, Ring A is substituted with a nitrile group or issubstituted at the para position with a 5- or 6-membered heterocycle.Typical examples of the heterocycle include pyrrolidyl, piperidinyl andmorpholinyl.

In yet another aspect, the invention provides novel sirtuin-modulatingcompounds of Formula (IV):Ar-L-J-M-K—Ar′  (IV)

-   -   or a salt thereof, wherein:    -   each Ar and Ar′ is independently an optionally substituted        carbocyclic or heterocyclic aryl group;    -   L is an optionally substituted carbocyclic or heterocyclic        arylene group;    -   each J and K is independently NR₁′, O, S, or is optionally        independently absent; or when J is NR₁′, R₁′ is a C1-C4 alkylene        or C2-C4 alkenylene attached to Ar′ to form a ring fused to Ar′;        or when K is NR₁′, R₁′ is a C1-C4 alkylene or C2-C4 alkenylene        attached to L to form a ring fused to L;    -   each M is C(O), S(O), S(O)₂, or CR₁′R₁′;    -   each R₁′ is independently selected from H, C1-C10 alkyl; C2-C10        alkenyl; C2-C10 alkynyl; C3-C10 cycloalkyl; C4-C10 cycloalkenyl;        aryl; R₅′; halo; haloalkyl; CF₃; SR₂′; OR₂′; NR₂′R₂′; NR₂′R₃′;        COOR₂′; NO₂; CN; C(O)R₂′; C(O)C(O)R₂′; C(O)NR₂′R₂′; OC(O)R₂′;        S(O)₂R₂′; S(O)₂NR₂′R₂′; NR₂′C(O)NR₂′R₂′; NR₂′C(O)C(O)R₂′;        NR₂′C(O)R₂′; NR₂′(COOR₂′); NR₂′C(O)R₅′; NR₂′S(O)₂NR₂′R₂′;        NR₂′S(O)₂R₂′; NR₂′S(O)₂R₅′; NR₂′C(O)C(O)NR₂′R₂′;        NR₂′C(O)C(O)NR₂′R₃′; C1-C10 alkyl substituted with aryl, R₄′ or        R₅′; or C2-C10 alkenyl substituted with aryl, R₄′ or R₅′;    -   each R₂′ is independently H; C1-C10 alkyl; C2-C10 alkenyl;        C2-C10 alkynyl; C3-C10 cycloalkyl; C4-C10 cycloalkenyl; aryl;        R₆′; C1-C10 alkyl substituted with 1-3 independent aryl, R₄′ or        R₆′ groups; C3-C10 cycloalkyl substituted with 1-3 independent        aryl, R₄′ or R₆′ groups; or C2-C10 alkenyl substituted with 1-3        independent aryl, R₄′ or R₆′;    -   each R₃′ is independently C(O)R₂′, COOR₂′, or S(O)₂R₂′;    -   each R₄′ is independently halo, CF₃, SR₇′, OR₇′, OC(O)R₇′,        NR₇′R₇′, NR₇′R₈′, NR₈′R₈′, COOR₇′, NO₂, CN, C(O)R₇′, or        C(O)NR₇′R₇′;    -   each R₅′ is independently a 5-8 membered monocyclic, 8-12        membered bicyclic, or 11-14 membered tricyclic ring system        comprising 1-3 heteroatoms if monocyclic, 1-6 heteroatoms if        bicyclic, or 1-9 heteroatoms if tricyclic, said heteroatoms        selected from O, N, or S, which may be saturated or unsaturated,        and wherein 0, 1, 2 or 3 atoms of each ring may be substituted        by a substituent independently selected from C1-C10 alkyl;        C2-C10 alkenyl; C2-C10 alkynyl; C3-C10 cycloalkyl; C4-C10        cycloalkenyl; aryl; R₆′; halo; sulfur; oxygen; CF₃; haloalkyl;        SR₂′; OR₂′; OC(O)R₂′; NR₂′R₂′; NR₂′R₃′; NR₃′R₃′; COOR₂′; NO₂;        CN; C(O)R₂′; C(O)NR₂′R₂′; C1-C10 alkyl substituted with 1-3        independent R₄′, R₆′, or aryl; or C2-C10 alkenyl substituted        with 1-3 independent R₄′, R₆′, or aryl;    -   each R₆′ is independently a 5-8 membered monocyclic, 8-12        membered bicyclic, or 11-14 membered tricyclic ring system        comprising 1-3 heteroatoms if monocyclic, 1-6 heteroatoms if        bicyclic, or 1-9 heteroatoms if tricyclic, said heteroatoms        selected from O, N, or S, which may be saturated or unsaturated,        and wherein 0, 1, 2 or 3 atoms of each ring may be substituted        by a substituent independently selected from C1-C10 alkyl;        C2-C10 alkenyl; C2-C10 alkynyl; C3-C10 cycloalkyl; C4-C10        cycloalkenyl; halo; sulfur; oxygen; CF₃; haloalkyl; SR₇′; OR₇′;        NR₇′R₇′; NR₇′R₈′; NR₈′R₈′; COOR₇′; NO₂; CN; C(O)R₇′; or        C(O)NR₇′R₇′;

each R₇′ is independently H, C1-C10 alkyl; C2-C10 alkenyl; C2-C10alkynyl; C3-C10 cycloalkyl; C4-C10 cycloalkenyl; haloalkyl; C1-C10 alkyloptionally substituted with 1-3 independent C1-C10 alkyl, C2-C10alkenyl, C2-C10 alkynyl, C3-C10 cycloalkyl, C4-C10 cycloalkenyl, halo,CF₃, OR₁₀′, SR₁₀′, NR₁₀′R₁₀′, COOR₁₀′, NO₂, CN, C(O)R₁₀′, C(O)NR₁₀′R₁₀′,NHC(O)R₁₀′, or OC(O)R₁₀′; or phenyl optionally substituted with 1-3independent C1-C10 alkyl, C2-C10 alkenyl, C2-C10 alkynyl, C3-C10cycloalkyl, C4-C10 cycloalkenyl, halo, CF₃, OR₁₀′, SR₁₀′, NR₁₀′R₁₀′,COOR₁₀′, NO₂, CN, C(O)R₁₀′, C(O)NR₁₀′R₁₀′, NHC(O)R₁₀′, or OC(O)R₁₀′;

-   -   each R₈′ is independently C(O)R₇′, COOR₇′, or S(O)₂R₇′;    -   each R₉′ is independently H, C1-C10 alkyl, C2-C10 alkenyl,        C2-C10 alkynyl, C3-C10 cycloalkyl, C4-C10 cycloalkenyl, or        phenyl optionally substituted with 1-3 independent C1-C10 alkyl,        C2-C10 alkenyl, C2-C10 alkynyl, C3-C10 cycloalkyl, C4-C10        cycloalkenyl, halo, CF₃, OR₁₀′, SR₁₀′, NR₁₀′R₁′, COOR₁₀′, NO₂,        CN, C(O)R₁₀′, C(O)NR₁₀′R₁₀′, NHC(O)R₁₀′, or OC(O)R₁₀′;    -   each R₁₀′ is independently H; C1-C10 alkyl; C2-C10 alkenyl;        C2-C10 alkynyl; C3-C10 cycloalkyl; C4-C10 cycloalkenyl; C1-C10        alkyl optionally substituted with halo, CF₃, OR₁₁′, SR₁₁′,        NR₁₁′R₁₁′, COOR₁₁′, NO₂, CN; or phenyl optionally substituted        with halo, CF₃, OR₁₁′, SR₁₁′, NR₁₁′R₁₁′, COOR₁′, NO₂, CN;    -   each R₁₁′ is independently H; C1-C10 alkyl; C3-C10 cycloalkyl or        phenyl;    -   each haloalkyl is independently a C1-C10 alkyl substituted with        one or more halogen atoms, selected from F, Cl, Br, or I,        wherein the number of halogen atoms may not exceed that number        that results in a perhaloalkyl group; and    -   each aryl is independently optionally substituted with 1-3        independent C1-C10 alkyl; C2-C10 alkenyl; C2-C10 alkynyl; C3-C10        cycloalkyl; C4-C10 cycloalkenyl; R₆′; halo; haloalkyl; CF₃;        OR₉′; SR₉′; NR₉′R₉′; COOR₉′; NO₂; CN; C(O)R₉′; C(O)C(O)R₉′;        C(O)NR₉′R₉′; S(O)₂R₉′; N(R₉′)C(O)R₉′; N(R₉′)(COOR₉′);        N(R₉′)S(O)₂R₉′; S(O)₂NR₉′R₉′; OC(O)R₉′; NR₉′C(O)NR₉′R₉′;        NR₉′C(O)C(O)R₉′; NR₉′C(O)R₆′; NR₉′S(O)₂NR₉′R₉′; NR₉′S(O)₂R₆′;        NR₉′C(O)C(O)NR₉′R₉′; C1-C10 alkyl substituted with 1-3        independent R₆′, halo, CF₃, OR₉′, SR₉′, NR₉′R₉′, COOR₉′, NO₂,        CN, C(O)R₉′, C(O)NR₉′R₉′, NHC(O)R₉′, NH(COOR₉′), S(O)₂NR₉′R₉′,        OC(O)R₉′; C2-C10 alkenyl substituted with 1-3 independent R₆′,        halo, CF₃, OR₉′, SR₉′, NR₉′R₉′, COOR₉′, NO₂, CN, C(O)R₉′,        C(O)NR₉′R₉′, NHC(O)R₉′, NH(COOR₉′), S(O)₂NR₉′R₉′, OC(O)R₉′; or        R₉′.

In a preferred embodiment of the invention, each Ar, L, and Ar′ isindependently an optionally substituted 5-to 7-membered monocyclic ringsystem or an optionally substituted 9-to 12-membered bicyclic ringsystem.

According to another preferred embodiment,

-   -   Ar is

-   -   X₁, X₂, X₃, X₄, and X₅ are independently selected from CR₁′ and        N; and    -   X₆ is selected from NR₁′, O, and S;

According to yet another preferred embodiment, X₁ and X₂ are N; X₃, X₄,and X₅ are CR₁′; and X₆ is O.

According to still yet another preferred embodiment, X₁ and X₃ are N;X₂, X₄, and X₅ are CR₁′; and X₆ is O.

According to still yet another preferred embodiment, X₁ and X₄ are N;X₂, X₃, and X₅ are CR₁′; and X₆ is O.

According to still yet another preferred embodiment, X₁ and X₅ are N;X₂, X₃, and X₄ are CR₁′; and X₆ is O.

In another embodiment, the compounds of the formula above are thosewherein J is NR₁′, K is absent, and M is C(O).

In yet another embodiment, the compounds of the formula above are thosewherein J is absent, K is NR₁′, and M is C(O).

In a further embodiment, compounds of formula (IV) are those where whenJ is absent and K is NR₁′, M is not C(O) and when J is NR₁′ and K isabsent, M is not C(O).

In a preferred embodiment, the compounds above are those wherein L is anoptionally substituted 5-to 7-membered carbocyclic or heterocyclic arylgroup.

In yet another preferred embodiment, the compounds are those wherein Lis an optionally substituted phenylene, pyridinylene, imidazolylene,oxazolylene, or thiazolylene.

In a particularly preferred embodiment, L is an optionally substitutedphenylene.

In another particularly preferred embodiment, L is an optionallysubstituted pyridinylene.

In an even more preferred embodiment, L is phenylene.

In another even more preferred embodiment, L is pyridinylene.

In either of these embodiments, Ar and J may be attached to L at theortho-, meta-, or para-positions. Particularly preferred are thoseembodiments where attachment is at the meta-position.

In certain embodiments, L is not phenylene when Ar′ is phenyl. Examplesof such embodiments include embodiments where L is an optionallysubstituted heterocyclic aryl group and Ar′ is an optionally substitutedcarbocyclic or heterocyclic aryl group, or wherein L is an optionallysubstituted carbocyclic or heterocyclic aryl group and Ar′ is anoptionally substituted heterocyclic aryl group.

In yet another aspect, the invention provides novel sirtuin-modulatingcompounds of Formula (I) or a salt thereof, wherein

-   -   Ring A is substituted with at least one R₁′ group;    -   R₁′, R₂′, R₃′, R₄′, R₅′, R₆′, R₇′, R₈′, R₉′, R₁₀′, and R₁₁′ are        as defined above;    -   each haloalkyl is independently a C1-C10 alkyl substituted with        one or more halogen atoms, selected from F, Cl, Br, or I,        wherein the number of halogen atoms may not exceed that number        that results in a perhaloalkyl group;    -   each aryl is independently a 5-to 7-membered monocyclic ring        system or a 9-to 12-membered bicyclic ring system optionally        substituted with 1-3 independent C1-C10 alkyl; C2-C10 alkenyl;        C2-C10 alkynyl; C3-C10 cycloalkyl; C4-C10 cycloalkenyl; R₆′;        halo; haloalkyl; CF₃; OR₉′; SR₉′; NR₉′R₉′; COOR₉′; NO₂; CN;        C(O)R₉′; C(O)C(O)R₉′; C(O)NR₉′R₉′; S(O)₂R₉′; N(R₉′)C(O)R₉′;        N(R₉′)(COOR₉′); N(R₉′)S(O)₂R₉′; S(O)₂NR₉′R₉′; OC(O)R₉′;        NR₉′C(O)NR₉′R₉′; NR₉′C(O)C(O)R₉′; NR₉′C(O)R₆′; NR₉′S(O)₂NR₉′R₉′;        NR₉′S(O)₂R₆′; NR₉′C(O)C(O)NR₉′R₉′; C1-C10 alkyl substituted with        1-3 independent R₆′, halo, CF₃, OR₉′, SR₉′, NR₉′R₉′, COOR₉′,        NO₂, CN, C(O)R₉′, C(O)NR₉′R₉′, NHC(O)R₉′, NH(COOR₉′),        S(O)₂NR₉′R₉′, OC(O)R₉′; C2-C10 alkenyl substituted with 1-3        independent R₆′, halo, CF₃, OR₉′, SR₉′, NR₉′R₉′, COOR₉′, NO₂,        CN, C(O)R₉′, C(O)NR₉′R₉′, NHC(O)R₉′, NH(COOR₉′), S(O)₂NR₉′R₉′,        OC(O)R₉′; or R₉′; and    -   Ring B is substituted with at least one

-   -    wherein    -   X₁, X₂, X₃, X₄, and X₅ are independently selected from CR₁′ and        N; and    -   X₆ is selected from NR₁′, O, and S.

In a preferred embodiment, Ring B is phenyl or pyridinyl.

In a further aspect, the invention provides novel sirtuin-modulatingcompounds of Formula (IVa):Het-L-Q-Ar′  (IVa).

-   or a salt thereof, wherein:    -   Het is an optionally substituted heterocyclic aryl group;    -   L is an optionally substituted carbocyclic or heterocyclic        arylene group;    -   Ar′ is an optionally substituted carbocyclic or heterocyclic        aryl group; and    -   Q is selected from —NR₁′—C(O)—, —NR₁′—S(O)₂—, —NR₁′—C(O)—NR₁′—,        —NR₁′—C(S)—NR₁′—, —NR₁′—C(O)—CR₁′R₁′—NR₁′—,        —NR₁′—C(═NR₁′)—NR₁′—, —C(O)—NR₁′—, —C(O)—NR₁′—S(O)₂—, —NR₁′—,        —CR₁′R₁′—, —NR₁′—C(O)—CR₁′═CR₁′—, —NR₁′—S(O)₂—NR₁′—,        —NR₁′—C(O)—NR₁′—S(O)₂—, —NR₁′—CR₁′R₁′—C(O)—NR₁′—,        —CR₁′R₁′—C(O)—NR₁′—, —NR₁′—C(O)—CR₁′═CR₁′—CR₁′R₁′—,        —NR₁′—C(═N—CN)—NR₁′—, —NR₁′—C(O)—CR₁′R₁′—,

-   -   each R₁′ is independently selected from H or optionally        substituted C₁-C₃ straight or branched alkyl, wherein:

-   when Het is a polycyclic heteroaryl, L is an optionally substituted    phenylene, Q and Het are attached to L in a meta orientation, and    Ar′ is optionally substituted phenyl; then Q is not —NH—C(O)—.

In certain embodiments, when Het is a polycyclic heteroaryl, L isoptionally substituted phenylene, and Ar′ is optionally substitutedphenyl; then Q is not —NH—C(O)—.

In certain embodiments (e.g., when the compound is a sirtuin activator),Het and Q are attached to L in a 1-, 2- or 1-,3-configuration (e.g.,when L is phenylene, Het and Q are attached in an ortho or a metaorientation). In certain embodiments where Het and Q are attached to Lin a 1-,3-configuration, if Het is benzoxazolyl, L is pyridylene and Qis —NH—C(O)—NH, then Ar′ is not 3,4 dioxymethylene phenyl; if Het ismethyl thiazolyl, L is phenylene and Q is —NH—C(O)—, then Ar′ is not3-dimethylamino phenyl; if Het is oxazolopyridyl, L is pyridylene and Qis —NH—C(O)—NH, then Ar′ is not 4-dimethylamino phenyl; if Het isoxazolopyridyl or benzoxazolyl and L is

then Q is not —NH—(SO)₂—; and if Het is oxazolopyridyl, L is

and Q is —NH—C(O)—, then Ar′ is not 3,4 dimethoxyphenyl or pyridyl.

When Het is substituted, it is typically substituted at up to 2 carbonatoms with a substituent independently selected from R₁₂, N(R₁₂)₂,NH(R₁₂), OR₁₂, C(O)—NH—R₁₂, C(O)—N(R₁₂)₂, N(R₁₂)—OR₁₂, CH₂—N(R₁₂)₂,C(O)OR₁₂, C(O)OH,

where each R₁₂ is independently selected from optionally substitutedC₁-C₃ straight or branched alkyl.

In certain embodiments, Het is selected from oxazolopyridyl,benzothienyl, benzofuryl, indolyl, quinoxalinyl, benzothiazolyl,benzoxazolyl, benzimidazolyl, quinolinyl, isoquinolinyl or isoindolyl.In other embodiments, Het comprises one ring N heteroatom and 1 to 2additional ring heteroatoms independently selected from N, O or S, suchas thiazolyl, triazolyl, oxadiazolyl, thiazolopyridyl, imidazothiazolyl,benzoxazinonyl, or imidazopyridyl.

Particular examples of Het include:

where up to 2 ring carbons not immediately adjacent to the indicatedattachment point are independently substituted with optionallysubstituted C₁-C₃ straight or branched alkyl, phenyl, halo, N(R₁₂)₂,NH(R₁₂), OR₁₂, C(O)—NH—R₁₂, C(O)—N(R₁₂)₂, N(R₁₂)—OR₁₂, CH₂—N(R₁₂)₂,C(O)OR₁₂, C(O)OH,

wherein each R₁₂ is independently selected from optionally substitutedC₁-C₃ straight or branched alkyl.

In certain embodiments, L is selected from

-   -   wherein:    -   each of Z₁, Z₂, Z₃ and Z₄ is independently selected from CH or        N, wherein not more than three of said Z₁, Z₂, Z₃ or Z₄ is N;    -   each of Z₅ and Z₆ is independently selected from C, Ng O or S,        provided that at least one of Z₅ and Z₆ is N; and    -   L is optionally substituted at 1 to 2 carbon atoms with a        substituent independently selected from R₁₂, N(R₁₂)₂, NH(R₁₂),        OR₁₂, C(O)—NH—R₁₂, C(O)—N(R₁₂)₂, N(R₁₂)—OR₁₂, CH₂—N(R₁₂)₂,        C(O)OR₁₂, C(O)OH,

In preferred embodiments, L is selected from phenylene or pyridylene,such as unsubstituted phenylene or phenylene substituted with a singlesubstituent selected from C(O)OCH₃, C(O)OH, CH₂OH, N(CH₃)₂, orCH₂N(CH₃)₂, or unsubstituted pyridylene.

In certain embodiments, Q is selected from —NH—C(O)—, —NH—S(O)₂—,—NH—C(O)—NH—, —C(O)—NH—, —CH₂—, —N(CH₃)—C(O)—NH—, —NH—C(O)—N(CH₃)—, or—NH—S(O)₂—NH—, particularly —NH—C(O)—, —C(O)—NH—, —NH—, —NH—C(O)—NH, or—NH—S(O)₂—.

In certain embodiments, Ar′ is selected from optionally substitutedphenyl, benzothiazolyl, or benzoxazolyl. When Ar′ is phenyl, typicaloptional substituents are 1 to 3 substituents independently selectedfrom halo, (optionally substituted C₁-C₃ straight or branched alkyl),O-(optionally substituted C₁-C₃ straight or branched alkyl),S-(optionally substituted C₁-C₃ straight or branched alkyl), N(CH₃)₂ oroptionally substituted heterocyclyl, or wherein two substituents onadjacent ring atoms are taken together to form a dioxymethylene.

In certain embodiments, Het is selected from

and wherein up to 2 ring carbons not immediately adjacent to theindicated attachment point are independently substituted with optionallysubstituted C₁-C₃ straight or branched alkyl, phenyl or halo;

-   -   L is selected from unsubstituted phenylene, phenylene        substituted with a single substituent selected from C(O)OCH₃,        C(O)OH, CH₂OH, N(CH₃)₂, or CH₂N(CH₃)₂, or unsubstituted        pyridylene;    -   Q is selected from —NH—C(O)—, —C(O)—NH—, —NH—, —NH—C(O)—NH, or        —NH—S(O)₂—; and    -   Ar′ is selected from optionally substituted phenyl,        benzothiazolyl, or benzoxazolyl, wherein said phenyl is        optionally substituted with 1 to 3 substituents independently        selected from chloro, methyl, O-methyl, S-methyl, N(CH₃)₂,        morpholino, or 3,4 dioxymethylene.

In certain embodiments, Q is selected from —NH—C(O)—, —C(O)—NH—, —NH— or—NH—C(O)—NH.

In certain embodiments, the substituents on Ar′ are selected fromchloro, methyl, O-methyl, S-methyl or N(CH₃)₂. In certain embodiments,the only substituent on Ar′ is an O-methyl group, particularly anO-methyl group ortho or meta to Q. In certain embodiments, when thereare two or more O-methyl groups or Ar′, at least one is ortho or meta toQ.

In certain embodiments, L is pyridyl and Het and Q are at the 1,3- or2,4-position with respect to the pyridyl nitrogen atom. In certain suchembodiments, Q is —NH—S(O)₂—.

In certain embodiments where L is further substituted, the substituentis typically meta to both Het and Q.

In certain embodiments, Q is —NH— and Het is thiazolyl oroxazolopyridyl.

In certain embodiments, Q is —NH— and Ar is benzothiazolyl orbenzoxazolyl.

In certain embodiments, such as when the sirtuin modulator is a sirtuinactivator, L is

and Q is —NH—(SO)₂—. In certain such embodiments, Het is oxazolopyridyl.When L, Q and optionally Het have these values, Ar′ is advantageouslynaphthyl or phenyl, where Ar′ is optionally substituted with 1 to 3substituents independently selected from CN, halo, (C₁-C₃ straight orbranched alkyl), O—(C₁-C₃ straight or branched alkyl), N(C₁-C₃ straightor branched alkyl)₂, or a 5 to 6-membered heterocycle.

In certain embodiments, such as when the sirtuin modulator is a sirtuinactivator, L is

and Q is —NH—C(O)—. In certain such embodiments, Het is oxazolopyridyl.When L, Q and optionally Het have these values, Ar′ is advantageouslypyridyl or phenyl optionally substituted with 1 to 3 substituentsindependently selected from CN, halo, (C₁-C₃ straight or branchedalkyl), O—(C₁-C₃ straight or branched alkyl), N(C₁-C₃ straight orbranched alkyl)₂, or a 5 to 6-membered heterocycle.

In certain embodiments, such as when the sirtuin modulatory is a sirtuininhibitor, Het comprises one N heteroatom and 1 to 2 additionalheteroatoms independently selected from N, O or S;

-   -   L is

-   -    and is optionally substituted;    -   Q is —NH—C(O)—; and    -   Ar′ is phenyl substituted with 1 to 3 substituents independently        selected from CN, halo, C₁-C₃ straight or branched alkyl,        O—(C₁-C₃ straight or branched alkyl), N(C₁-C₃ straight or        branched alkyl)₂, or a 5 to 6-membered heterocycle, wherein when        R₈ is unsubstituted

-   -    then ring A is:    -   a) not simultaneously substituted at the 2-and 6-positions with        O—(C₁-C₃ straight or branched alkyl);    -   b) not simultaneously substituted at the 2-position with C₁-C₃        straight or branched alkyl or O—(C₁-C₃ straight or branched        alkyl) and at the 3-position with O—(C₁-C₃ straight or branched        alkyl);    -   c) not substituted at the 4-position with O—(C₁-C₃ straight or        branched alkyl) unless simultaneously substituted at the        3-position with halo or O—(C₁-C₃ straight or branched alkyl) and        unsubstituted at all other positions;        not substituted at the 4-position with N(C₁-C₃ straight or        branched alkyl)₂, or said 5 to 6-membered heterocycle. In        certain such embodiments, L is unsubstituted and/or Het is        oxazolopyridyl.

In yet another aspect, the invention provides novel sirtuin-modulatingcompounds of Formula (V):

-   -   or a salt thereof, wherein:    -   Ring A is optionally substituted with at least one R₁′ group;    -   Y₁, Y₂, Y₃, Y₄, and Y₅ are independently R₁′;    -   R₁′, R₂′, R₃′, R₄′, R₅′, R₆′, R₇′, R₈′, R₉′, R₁₀′, and R₁₁′ are        as defined above;    -   each haloalkyl is independently a C1-C10 alkyl substituted with        one or more halogen atoms, selected from F, Cl, Br, or I,        wherein the number of halogen atoms may not exceed that number        that results in a perhaloalkyl group; and    -   each aryl is independently a 5-to 7-membered monocyclic ring        system or a 9-to 12-membered bicyclic ring system optionally        substituted with 1-3 independent C1-C10 alkyl; C2-C10 alkenyl;        C2-C10 alkynyl; C3-C10 cycloalkyl; C4-C10 cycloalkenyl; R₆′;        halo; haloalkyl; CF₃; OR₉′; SR₉′; NR₉′R₉′; COOR₉′; NO₂; CN;        C(O)R₉′; C(O)C(O)R₉′; C(O)NR₉′R₉′; S(O)₂R₉′; N(R₉′)C(O)R₉′;        N(R₉′)(COOR₉′); N(R₉′)S(O)₂R₉′; S(O)₂NR₉′R₉′; OC(O)R₉′;        NR₉′C(O)NR₉′R₉′; NR₉′C(O)C(O)R₉′; NR₉′C(O)R₆′; NR₉′S(O)₂NR₉′R₉′;        NR₉′S(O)₂R₆′; NR₉′C(O)C(O)NR₉′R₉′; C1-C10 alkyl substituted with        1-3 independent R₆′, halo, CF₃, OR₉′, SR₉′, NR₉′R₉′, COOR₉′,        NO₂, CN, C(O)R₉′, C(O)NR₉′R₉′, NHC(O)R₉′, NH(COOR₉′),        S(O)₂NR₉′R₉′, OC(O)R₉′; C2-C10 alkenyl substituted with 1-3        independent R₆′, halo, CF₃, OR₉′, SR₉′, NR₉′R₉′, COOR₉′, NO₂,        CN, C(O)R₉′, C(O)NR₁′R₉′, NHC(O)R₉′, NH(COOR₉′), S(O)₂NR₉′R₉′,        OC(O)R₉′; or R₉′.

In a preferred embodiment of the above compound,

-   -   either Y₂ or Y₃ is

-   -   X₁, X₂, X₃, X₄, and X₅ are independently selected from CR₁′ and        N; and    -   X₆ is selected from NR₁′, O, and S.

According to an even more preferred embodiment, X₁ and X₂ are N; X₃, X₄,and X₅ are CR₁′; and X₆ is O.

According to another even more preferred embodiment, X₁ and X₃ are N;X₂, X₄, and X₅ are CR₁′; and X₆ is O.

According to another even more preferred embodiment, X₁ and X₄ are N;X₂, X₃, and X₅ are CR₁′; and X₆ is O.

According to another even more preferred embodiment, X₁ and X₅ are N;X₂, X₃, and X₄ are CR₁′; and X₆ is O.

In another aspect, the invention provides sirtuin-modulating compoundsof Structural Formula (VII):

-   or a salt thereof, wherein:    -   each of X₇, X₈, X₉ and X₁₀ is independently selected from N,        CR²⁰, or CR₁′, wherein:        -   each R²⁰ is independently selected from H or a solubilizing            group;        -   each R₁′ is independently selected from H or optionally            substituted C₁-C₃ straight or branched alkyl;        -   one of X₇, X₈, X₉ and X₁₀ is N and the others are selected            from CR²⁰ or CR₁′; and        -   zero to one R²⁰ is a solubilizing group;    -   R¹⁹ is selected from:

-   -   wherein:        -   each Z₁₀, Z₁₀, Z₁₂ and Z₁₃ is independently selected from N,            CR²⁰, or CR₁′; and        -   each Z₁₄, Z₁₅ and Z₁₆ is independently selected from N,            NR₁′, S, O, CR²⁰, or CR₁′, wherein:        -   zero to two of Z₁₀, Z₁₁, Z₁₂ or Z₁₃ are N;        -   at least one of Z₁₄, Z₁₅ and Z₁₆ is N, NR₁′, S or O;        -   zero to one of Z₁₄, Z₁₅ and Z₁₆ is S or O;        -   zero to two of Z₁₄, Z₁₅ and Z₁₆ are N or NR₁′;        -   zero to one R²⁰ is a solubilizing group;        -   zero to one R₁′ is an optionally substituted C₁-C₃ straight            or branched alkyl; and    -   R²¹ is selected from —NR₁′—C(O)—, —NR₁′—S(O)₂—,        —NR₁′—C(O)—NR₁′—, —NR₁′—C(S)—NR₁′—, —NR₁′—C(S)—NR₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—NR₁′—, —NR₁′—C(═NR₁′)—NR₁′—, —C(O)—NR₁′—,        —C(O)—NR₁′—S(O)₂—, —NR₁′—, —CR₁′R₁′—, —NR₁′—C(O)—CR₁′═CR₁′—,        —NR₁′—S(O)₂—NR₁′—, —NR₁′—C(O)—NR₁′—S(O)₂—,        —NR₁′—CR₁′R₁′—C(O)—NR₁′—, —CR₁′R₁′—C(O)—NR₁′—,        —NR₁′—C(O)—CR₁′═CR₁′—CR₁′R₁′—, —NR₁′—C(═N—CN)—NR₁′—,        —NR₁′—C(O)—CR₁′R₁′—O—, —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—O—,        —NR₁′—S(O)₂—CR₁′R₁′—, —NR₁′—S(O)₂—CR₁′R₁′—CR₁′R₁′—, or        —NR₁′—C(O)—CR₁′R₁′—; and    -   R³¹ is selected from an optionally substituted monocyclic or        bicyclic aryl, or an optionally substituted monocyclic or        bicyclic heteroaryl, with the provisos that said compound is        not:

-   -    that when R¹⁹ is

-   -    and R²¹ is —NHC(O)—, R³¹ is not an optionally substituted        phenyl.

In certain embodiments, compounds of Structural Formula (VII) have thefollowing values:

-   -   each of X₇, X₈, X₉ and X₁₀ is independently selected from N,        CR²⁰, or CR₁′, wherein:    -   each R²⁰ is independently selected from H or a solubilizing        group;    -   each R₁′ is independently selected from H or optionally        substituted C₁-C₃ straight or branched alkyl;    -   one of X₇, X₈, X₉ and X₁₀ is N and the others are selected from        CR²⁰ or CR₁′; and    -   zero to one R²⁰ is a solubilizing group;    -   R¹⁹ is selected from:

-   -   wherein:        -   each Z₁₀, Z₁₁, Z₁₂ and Z₁₃ is independently selected from N,            CR²⁰, or CR₁′; and        -   each Z₁₄, Z₁₅ and Z₁₆ is independently selected from N,            NR₁′, S, O, CR²⁰, or CR₁′, wherein:        -   zero to two of Z₁₀, Z₁₁, Z₁₂ or Z₁₃ are N;        -   at least one of Z₁₄, Z₁₅ and Z₁₆ is N, NR₁′, S or O;        -   zero to one of Z₁₄, Z₁₅ and Z₁₆ is S or O;        -   zero to two of Z₁₄, Z₁₅ and Z₁₆ are N or NR₁′;        -   zero to one R²⁰ is a solubilizing group;        -   zero to one R₁′ is an optionally substituted C₁-C₃ straight            or branched alkyl; and    -   R²¹ is selected from —NR₁′—C(O)—, —NR₁′—S(O)₂—,        —NR₁′—C(O)—NR₁′—, —NR₁′—C(S)—NR₁′—, —NR₁′—C(S)—NR₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—NR₁′—, —NR₁′—C(═NR₁′)—NR₁′—, —C(O)—NR₁′—,        —C(O)—NR₁′—S(O)₂—, —NR₁′—, —CR₁′R₁′—, —NR₁′—C(O)—CR₁′═CR₁′—,        —NR₁′—S(O)₂—NR₁′—, —NR₁′—C(O)—NR₁′—S(O)₂—,        —NR₁′—CR₁′R₁′—C(O)—NR₁′—, —CR₁′R₁′—C(O)—NR₁′—,        —NR₁′—C(O)—CR₁′═CR₁′—CR₁′R₁′—, —NR₁′—C(═N—CN)—NR₁′—,        —NR₁′—C(O)—CR₁′R₁′—O—, —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—O—,        —NR₁′—S(O)₂—CR₁′R₁′—, —NR₁′—S(O)₂—CR₁′R₁′CR₁′R₁′—, or        —NR₁′—C(O)—CR₁′R₁′—; and    -   R³¹ is selected from an optionally substituted monocyclic or        bicyclic aryl, or an optionally substituted monocyclic or        bicyclic heteroaryl, with the provisos that:    -   said compound is not:

-   -   when X₈ and X₉ are each independently selected from CR²⁰ or        CR₁′, R¹⁹ is

-   -    and each of Z₁₀, Z₁₁, Z₁₂ and Z₁₃ is independently selected        from CR²⁰, or CR₁′, then:    -   a) at least one of X₈ and X₉ is not CH; or    -   b) at least one of Z₁₀, Z₁₁, Z₁₂ and Z₁₃ is CR²⁰, wherein R²⁰ is        a solubilizing group.

In certain embodiments, when Z₁₂ is CR²⁰ and R²⁰ is a solubilizinggroup, the solubilizing group is not —C(O)OCH₂CH₃, —COOH,

In certain embodiments, when X₈ and X₉ are each independently selectedfrom CR²⁰ or CR₁′, R¹⁹ is

and each of Z₁₀, Z₁₁, Z₁₂ and Z₁₃ is independently selected from CR²⁰,or CR₁′, then:

-   -   a) at least one of X₈ and X₉ is not CH; or    -   b) at least one of Z₁₀, Z₁₁ and Z₁₃ is CR²⁰, wherein R²⁰ is a        solubilizing group.

In certain embodiments, when R¹⁹ is

and each of Z₁₀, Z₁₁, Z₁₂ and Z₁₃ is CR²⁰, or CR₁′; X₈ and X₉ are CR²⁰or CR₁′; R²¹ is —NHC(O)—; and R³¹ is optionally substituted phenyl, thenR³¹ is a substituted phenyl, at least one R₁′ in a CR₁′ moiety isoptionally substituted C₁-C₃ straight or branched alkyl, or at least oneR²⁰ in a CR²⁰ is a solubilizing group, or a combination thereof.

In certain embodiments, R¹⁹ is selected from phenyl, pyridyl, thienyl orfuryl.

In certain embodiments, R¹⁹ is

wherein each of Z₁₀, Z₁₁, Z₁₂ and Z₁₃ is independently selected fromCR²⁰ or CR₁′; and

-   -   R²¹ is —NH—C(O)—; and    -   R³¹ is a substituted phenyl.        In certain such embodiments, when X₉ is N, R³¹ is not 2,4        dimethoxyphenyl and/or when X₁₀ is N, R³¹ is not halo        substituted phenyl; 3,4-dioxoethylenephenyl; or        3,5-dimethoxyphenyl.

In preferred embodiments, R³¹ is optionally substituted with 1 to 3substituents independently selected from —OCH₃, —CH₃, —N(CH₃)₂,pyrazinoxy or a solubilizing group. Suitable examples of R³¹ include3-methoxy-4-((4-methylpiperazin-1-yl)methyl)phenyl,3-methoxy-4-morpholinomethylphenyl, 3-methoxy-4-diaminomethylphenyl,3-methoxy-4-((pyrrolidin-1-yl)methyl)phenyl, 3,4-dimethoxyphenyl,3,5-dimethoxyphenyl, 2,3,4-trimethoxyphenyl, 3,4,5-trimethoxyphenyl,2-dimethylaminophenyl, 3-dimethylaminophenyl, 4-dimethylaminophenyl, or3,5-dimethylphenyl.

In certain embodiments, R¹⁹ is selected from

wherein one of Z₁₀, Z₁₁, Z₁₂, and Z₁₃ is N and the others areindependently selected from CR²⁰ or CR₁′;

-   -   R²¹ is selected from —NH—, —NH—C(O)—, —NH—C(O)—NH, —NH—C(S)—NH—        or —NH—S(O)₂—; and    -   R³¹ is selected from an optionally substituted phenyl, an        optionally substituted naphthyl, or an optionally substituted        heteroaryl.

In certain such embodiments,

-   -   a) when R²¹ is —NH—S(O)₂—, either:        -   i) Z₁₀ is N; or        -   ii) Z₁₁ is N and R³¹ is halophenyl or            2-methoxy-5-methylphenyl;    -   b) when R³⁹ is

-   -    R³¹ is not 4-dimethylaminophenyl, 2,3,4-trimethoxyphenyl, or        3,5 dimethoxyphenyl; and/or    -   c) when R²¹ is —NH—C(O)—NH— and Z₁₀ is N, R³¹ is not        4-dimethylaminophenyl.

In certain such embodiments, R³¹ is selected from optionally substitutedphenyl, benzothiazolyl, or benzoxazolyl.

In yet another embodiment, the invention provides sirtuin-modulatingcompounds of Structural Formula (VIII):

-   or a salt thereof, wherein:    -   R₁′ is selected from H or optionally substituted C₁-C₃ straight        or branched alkyl;    -   R²¹ is selected from —NR₁′—C(O)—, —NR₁′—S(O)₂—,        —NR₁′—C(O)—NR₁′—, —NR₁′—C(S)—NR₁′—, —NR₁′—C(S)—NR₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—NR₁′—, —NR₁′—C(═NR₁′)—NR₁′—, —C(O)—NR₁′—,        —C(O)—NR₁′—S(O)₂—, —NR₁′—, —CR₁′R₁′—, —NR₁′—C(O)—CR₁′═CR₁′—,        —NR₁′—S(O)₂—NR₁′—, —NR₁′—C(O)—NR₁′—S(O)₂—,        —NR₁′—CR₁′R₁′—C(O)—NR₁′—, —CR₁′R₁′—C(O)—NR₁′—,        —NR₁′—C(O)—CR₁′═CR₁′—CR₁′R₁′—, —NR₁′—C(═N—CN)—NR₁′—,        —NR₁′—C(O)—CR₁′R₁′—O—, —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—O—,        —NR₁′—S(O)₂—CR₁′R₁′—, —NR₁′—S(O)₂—CR₁′R₁′—CR₁′R₁′—, or        —NR₁′—C(O)—CR₁′R₁′—; and    -   R³¹ is selected from an optionally substituted monocyclic or        bicyclic aryl, or an optionally substituted monocyclic or        bicyclic heteroaryl, with the provisos that:    -   when R₁′ is methyl, and R²¹ is —NH—C(O)—, R³¹ is not

-   -    1-methoxynaphthyl, 2-methoxynaphthyl, or unsubstituted        2-thienyl;    -   when R₁′ is methyl, and R²¹ is —NH—C(O)—CH═CH—, R³¹ is not

-   -   when R₁′ is methyl, and R²¹ is —NH—C(O)—CH—O—, R³¹ is not        unsubstituted naphthyl, 2-methoxy, 4-nitrophenyl,        4-chloro-2-methylphenyl, or 4-t-butylphenyl; and    -   when R²¹ is —NH—C(O)—, R³¹ is not optionally substituted phenyl.

In certain embodiments, R²¹ is —NH—C(O)—; and R³¹ is phenyl optionallysubstituted with 1 to 3 substituents independently selected from —OCH₃,—CH₃, —N(CH₃)₂, or a solubilizing group.

In certain such embodiments, R²¹ is —NH—C(O)— and R³¹ is selected fromunsubstituted phenyl, 2-methoxyphenyl, 3-methoxyphenyl,2,3,4-trimethoxyphenyl, 3,4,5-trimethoxyphenyl, 2,4-dimethoxyphenyl,3,5-dimethoxyphenyl, 2-methyl-3-methoxyphenyl, 2-morpholinophenyl,2-methoxy-4-methylphenyl, 2-dimethylaminophenyl, 4-dimethylaminophenyl,or

particularly phenyl; 2-methoxyphenyl; 3-methoxyphenyl;2,3,4-trimethoxyphenyl; 3,4,5-trimethoxyphenyl; 2,4-dimethoxyphenyl;3,5-dimethoxyphenyl; 2-methyl-3-methoxyphenyl; 2-morpholinophenyl;2-methoxy-4-methylphenyl; 2-dimethylaminophenyl; or4-dimethylaminophenyl.

In a further embodiment, the invention provides sirtuin-modulatingcompounds of Structural Formula (IX):

-   or a salt thereof, wherein:    -   R₁′ is selected from H or optionally substituted C₁-C₃ straight        or branched alkyl; and    -   R⁵⁰ is selected from 2,3-dimethoxyphenyl, phenoxyphenyl,        2-methyl-3-methoxyphenyl, 2-methoxy-4-methylphenyl, or phenyl        substituted with 1 to 3 substituents, wherein one of said        substituents is a solubilizing group; with the provisos that R⁵⁰        is not substituted simultaneously with a solubilizing group and        a nitro group, and R⁵⁰ is not singly substituted at the        4-position with cyclic solubilizing group or at the 2-position        with a morpholino group.

In one aspect, the invention provides sirtuin-modulating compounds ofStructural Formula (X):

-   or a salt thereof, wherein:    -   R₁′ is selected from H or optionally substituted C₁-C₃ straight        or branched alkyl; and    -   R⁵¹ is selected from an optionally substituted monocyclic        heteroaryl, an optionally substituted bicyclic heteroaryl, or an        optionally substituted naphthyl, wherein R⁵¹ is not        chloro-benzo[b]thienyl, unsubstituted benzodioxolyl,        unsubstituted benzofuranyl, methyl-benzofuranyl, unsubstituted        furanyl, phenyl-, bromo-, or nitro-furyl,        chlorophenyl-isoxazolyl, oxobenzopyranyl, unsubstituted        naphthyl, methoxy-, methyl-, or halo-naphthyl, unsubstituted        thienyl, unsubstituted pyridinyl, or chloropyridinyl.

In certain embodiments, R⁵¹ is selected from pyrazolyl, thiazolyl,oxazolyl, pyrimidinyl, furyl, thienyl, pyridyl, isoxazolyl, indolyl,benzopyrazolyl, benzothiazolyl, benzoxazolyl, quinoxalinyl,benzofuranyl, benzothienyl, quinolinyl, benzoisoxazolyl, benzotriazinyl,triazinyl, naphthyl, or

and wherein R⁵¹ is optionally substituted. In certain such embodiments,R⁵¹ is selected from pyrazolyl, thiazolyl, oxazolyl, pyrimidinyl,indolyl, pyrazinyl, triazinyl, or

and R⁵¹ is optionally substituted.

In another aspect, the invention provides sirtuin-modulating compoundsof Structural Formula (XI):

-   or a salt thereof, wherein:    -   R₁′ is selected from H or optionally substituted C₁-C₃ straight        or branched alkyl;    -   R²² is selected from —NR²³—C(O)—, —NR₁′—S(O)₂—,        —NR₁′—C(O)—NR₁′—, —NR₁′—C(S)—NR₁′—, —NR₁′—C(S)—NR₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—NR₁′—, —NR₁′—C(═NR₁′)—NR₁′—, —C(O)—NR₁′—,        —C(O)—NR₁′—S(O)₂—, —NR₁′—, —CR₁′R₁′—, —NR₁′—C(O)—CR₁′═CR₁′—,        —NR₁′—S(O)₂—NR₁′—, —NR₁′—C(O)—NR₁′—S(O)₂—,        —NR₁′—CR₁′R₁′—C(O)—NR₁′—, —CR₁′R₁′—C(O)—NR₁′—,        —NR₁′—C(O)—CR₁′═CR₁′—CR₁′R₁′—, —NR₁′—C(═N—CN)—NR₁′—,        —NR₁′—C(O)—CR₁′R₁′—O—, —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—O—,        —NR₁′—S(O)₂—CR₁′R₁′—, —NR₁′—S(O)₂—CR₁′R₁′—CR₁′R₁′—, or        —NR₁′—C(O)—CR₁′R₁′—, wherein R²³ is an optionally substituted        C₁-C₃ straight or branched alkyl; and    -   R³¹ is selected from an optionally substituted monocyclic or        bicyclic aryl, or an optionally substituted monocyclic or        bicyclic heteroaryl, with the provisos that:    -   when R²² is —NH—C(O)—CH═CH—, R³¹ is not unsubstituted furyl,        5-(2-methyl-3-chlorophenyl)-furanyl, 2,4-dichlorophenyl,        3,5-dichloro-2-methoxyphenyl, 3-nitrophenyl, 4-chlorophenyl,        4-chloro-3-nitrophenyl, 4-isopropylphenyl, 4-methoxyphenyl,        2-methoxy-5-bromophenyl, or unsubstituted phenyl;    -   when R²² is —NH—C(O)—CH₂—, R³¹ is not 3,4-dimethoxyphenyl,        4-chlorophenyl, or unsubstituted phenyl;    -   when R²² is —NH—C(O)—CH₂—O—, R³¹ is not        2,4-dimethyl-6-nitrophenyl, 2- or 4-nitrophenyl,        4-cyclohexylphenyl, 4-methoxyphenyl, unsubstituted naphthyl, or        unsubstituted phenyl, or phenyl mono substituted, disubstituted        or trisubstituted solely with substituents selected from        straight- or branched-chain alkyl or halo;    -   when R²² is —NH—C(O)—CH(CH₃)—O—, R³¹ is not 2,4-dichlorophenyl,        4-chlorophenyl, or unsubstituted phenyl; and    -   when R²² is —NH—S(O)₂—, R³¹ is not unsubstituted phenyl.

In certain embodiments, R²² is selected from —C(O)—NH—, —NH—, or—C(O)—NH—CH₃.

In certain embodiments, such as when R²² is selected from —C(O)—NH—,—NH—, or —C(O)—NH—CH₃, R³¹ is selected from optionally substitutedphenyl, benzothiazolyl, quinoxalinyl, or benzoxazolyl.

In yet another aspect, the invention provides sirtuin-modulatingcompounds of Structural Formula (XII):

-   or a salt thereof, wherein:    -   each of X₇, X₈, X₉ and X₁₀ is independently selected from N,        CR²⁰, or CR₁′, wherein:        -   each R²⁰ is independently selected from H or a solubilizing            group;        -   each R₁′ is independently selected from H or optionally            substituted C₁-C₃ straight or branched alkyl;        -   one of X₇, X₈, X₉ and X₁₀ is N and the others are selected            from CR²⁰ or CR₁′; and        -   zero to one R²⁰ is a solubilizing group;    -   R¹⁹ is selected from:

-   -   wherein:        -   each Z₁₀, Z₁₁, Z₁₂ and Z₁₃ is independently selected from N,            CR²⁰, or CR₁′; and        -   each Z₁₄, Z₁₅ and Z₁₆ is independently selected from N,            NR₁′, S, O, CR²⁰, or CR₁′, wherein:        -   zero to two of Z₁₀, Z₁₁, Z₁₂ or Z₁₃ are N;        -   at least one of Z₁₄, Z₁₅ and Z₁₆ is N, NR₁′, O or S;        -   zero to one of Z₁₄, Z₁₅ and Z₁₆ is S or O;        -   zero to two of Z₁₄, Z₁₅ and Z₁₆ are N or NR₁′;        -   zero to one R²⁰ is a solubilizing group;        -   zero to one R₁′ is an optionally substituted C₁-C₃ straight            or branched alkyl; and    -   R²¹ is selected from —NR₁′—C(O)—, —NR₁′—S(O)₂—,        —NR₁′—C(O)—NR₁′—, —NR₁′—C(S)—NR₁′—, —NR₁′—C(S)—NR₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—NR₁′—, —NR₁′—C(═NR₁′)—NR₁′—, —C(O)—NR₁′—,        —C(O)—NR₁′—S(O)₂—, —NR₁′—, —CR₁′R₁′—, —NR₁′—C(O)—CR₁′—CR₁′—,        —NR₁′—S(O)₂—NR₁′—, —NR₁′—C(O)—NR₁′—S(O)₂—,        —NR₁′—CR₁′R₁′—C(O)—NR₁′—, —CR₁′R₁′—C(O)—NR₁′—,        —NR₁′—C(O)—CR₁′═CR₁′—CR₁′R₁′—, —NR₁′—C(═N—CN)—NR₁′—,        —NR₁′—C(O)—CR₁′R₁′—O—, —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—O—,        —NR₁′—S(O)₂—CR₁′R₁′—, —NR₁′—S(O)₂—CR₁′R₁′—CR₁′R₁′—, or        —NR₁′—C(O)—CR₁′R₁′—; and    -   R³¹ is selected from an optionally substituted monocyclic or        bicyclic aryl, or an optionally substituted monocyclic or        bicyclic heteroaryl, with the proviso that when R¹⁹ is

-   -    Z₁₀, Z₁₁, Z₁₂ and Z₁₃ are each CH, and R²¹ is —NHC(O)—, R³¹ is        not an optionally substituted phenyl.

In certain embodiments, the compounds of Structural Formula (XI) havethe following values:

-   -   each of X₇, X₈, X₉ and X₁₀ is independently selected from N,        CR²⁰, or CR₁′, wherein:        -   each R²⁰ is independently selected from H or a solubilizing            group;        -   each R₁′ is independently selected from H or optionally            substituted C₁-C₃ straight or branched alkyl;        -   one of X₇, X₈, X₉ and X₁₀ is N and the others are selected            from CR²⁰ or CR₁′; and        -   zero to one R²⁰ is a solubilizing group;    -   R¹⁹ is selected from:

-   -   wherein:        -   each Z₁₀, Z₁₁, Z₁₂ and Z₁₃ is independently selected from N,            CR²⁰, or CR₁′; and        -   each Z₁₄, Z₁₅ and Z₁₆ is independently selected from N,            NR₁′, S, O, CR²⁰, or CR₁′, wherein:        -   zero to two of Z₁₀, Z₁₁, Z₁₂ or Z₁₃ are N;        -   at least one of Z₁₄, Z₁₅ and Z₁₆ is N, NR₁′, S or O;        -   zero to one of Z₁₄, Z₁₅ and Z₁₆ is S or O;        -   zero to two of Z₁₄, Z₁₅ and Z₁₆ are N or NR₁′;        -   zero to one R²⁰ is a solubilizing group;        -   zero to one R₁′ is an optionally substituted C₁-C₃ straight            or branched alkyl; and    -   R²¹ is selected from —NR₁′—C(O)—, —NR₁′—S(O)₂—,        —NR₁′—C(O)—NR₁′—, —NR₁′—C(S)—NR₁′—, —NR₁′—C(S)—NR₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—NR₁′—, —NR₁′—C(═NR₁′)—NR₁′—, —C(O)—NR₁′—,        —C(O)—NR₁′—S(O)₂—, —NR₁′—, —CR₁′R₁′—, —NR₁′—C(O)—CR₁′═CR₁′—,        —NR₁′—S(O)₂—NR₁′—, —NR₁′—C(O)—NR₁′—S(O)₂—,        —NR₁′—CR₁′R₁′—C(O)—NR₁′—, —CR₁′R₁′—C(O)—NR₁′—,        —NR₁′—C(O)—CR₁′═CR₁′—CR₁′R₁′—, —NR₁′—C(═N—CN)—NR₁′—,        —NR₁′—C(O)—CR₁′R₁′—O—, —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—O—,        —NR₁′—S(O)₂—CR₁′R₁′—, —NR₁′—S(O)₂—CR₁′R₁′—CR₁′R₁′—, or        —NR₁′—C(O)—CR₁′R₁′—; and    -   R³¹ is selected from an optionally substituted monocyclic or        bicyclic aryl, or an optionally substituted monocyclic or        bicyclic heteroaryl, with the proviso that:    -   when X₇ is N, R¹⁹ is

-   -    and each of Z₁₀, Z₁₁, Z₁₂ and Z₁₃ is independently selected        from CR²⁰, or CR₁′, then:        -   a) at least one of X₈, X₉ or X₁₀ is C—(C₁-C₃ straight or            branched alkyl) or C-(solubilizing group); or        -   b) at least one of Z₁₀, Z₁₁, Z₁₂ and Z₁₃ is CR²⁰, wherein            R²⁰ is a solubilizing group.

In certain embodiments, R²¹ is —NH—C(O)— and R¹⁹ is selected from:

In certain embodiments, R¹⁹ is selected from optionally substitutedphenyl, optionally substituted pyridyl, optionally substituted thienylor optionally substituted furyl.

In certain embodiments, R¹⁹ is

wherein each of Z₁₀, Z₁₁, Z₁₂ and Z₁₃ is independently selected fromCR²⁰ or CR₁′; and

-   -   R²¹ is selected from —NH—C(O)—, —NH—C(O)—CH(CH₃)—O—,        —NH—C(O)—CH₂—O—, or —NH—S(O)₂—CH₂—CH₂—; and    -   R³¹ is selected from an optionally substituted aryl, or an        optionally substituted heteroaryl.

In certain such embodiments, R³¹ is optionally substituted with 1 to 3substituents independently selected from —OCH₃, —CH₃, —N(CH₃)₂, phenyl,phenoxy, 3,4-dioxymethylene, fluoro, or another solubilizing group.Suitable examples of R³¹ include unsubstituted quinolinyl,2,4-dimethoxyphenyl, 3,4-dimethoxyphenyl, 3,5-dimethoxyphenyl,3,4,5-trimethoxyphenyl, 2,3,4-trimethoxyphenyl, 2-dimethylaminophenyl,3-dimethylaminophenyl, 4-dimethylaminophenyl, 3,5-dimethylphenyl,3,5-difluorophenyl, 3-trifluoromethoxyphenyl, unsubstitutedquinoxalinyl, unsubstituted benzopyrimidinyl,

In certain such embodiments, R³¹ is not phenyl-substituted furyl.

In certain embodiments, R¹⁹ is selected from

-   -   each of Z₁₀, Z₁₁, Z₁₂ and Z₁₃ is independently selected from        CR²⁰, or CR₁′;    -   R²¹ is selected from —NH—C(O)—, NH—C(O)—CH₂—CH(CH₃)—O,        —NH—C(O)—NH—, —NH—C(S)—NH—, —NH—C(S)—NH—CH₂—, or —NH—S(O)₂—; and    -   R³¹ is selected from an optionally substituted phenyl, an        optionally substituted naphthyl, or an optionally substituted        heteroaryl.

In certain such embodiments, R³¹ is selected from phenyl, naphthyl,pyrazolyl, furyl, thienyl, pyridyl, isoxazolyl, benzopyrazolyl,benzofuryl, benzothienyl, quinolinyl, benzoisoxazolyl, or

and R³¹ is optionally substituted (e.g., optionally substituted with upto three substituents independently selected from —OCH₃, —CH₃, —N(CH₃)₂,—O-phenyl, or another solubilizing group). Suitable examples of R³¹include unsubstituted phenyl, 3-methoxyphenyl, 4-methoxyphenyl, 2,3dimethoxyphenyl, 2,4-dimethoxyphenyl, 2,5-bis(trifluoromethyl)phenyl,3,4-dimethoxyphenyl, 3,5-dimethoxyphenyl, 3,4,5-trimethoxyphenyl,2,3,4-trimethoxyphenyl, 2-methoxy-4-methylphenyl, 2-phenoxyphenyl,3-dimethylaminophenyl, 4-dimethylaminophenyl, unsubstituted 2-furanyl,unsubstituted 2-thienyl,

In certain embodiments, one or more of the following conditions applies:

-   -   when X₈ is N, R²¹ is —NH—C(S)—NH—, and R¹⁹ is phenyl, R³¹ is not        2-methoxy-5-nitrophenyl, 2-S-methylphenyl or 2-acetylphenyl;    -   when X₈ is N, R²¹ is —NH—S(O)₂—, and R¹⁹ is phenyl, R³¹ is not        thiadiazole-substituted thienyl or 4-methylsulfonylphenyl;    -   when X₈ is N, R²¹ is —NH—CO—, and R¹⁹ is phenyl, R³¹ is not        2,4-difluorophenyl, pyridyl-substituted thienyl,        3,4-dichlorophenyl, 4-t-butylphenyl, or 3-benzyloxyphenyl;    -   when X₉ is N, R²¹ is —NH—C(O)— and R¹⁹ is

-   -    R³¹ is not 2,3,4-trimethoxyphenyl or 3,5-dimethoxyphenyl; and    -   when X₉ is N, R²¹ is —NH—C(O)— and R¹⁹ is phenyl, R³¹ is not        3,5-dimethoxyphenyl.

In a further embodiment, the invention provides compounds of StructuralFormula (XIII):

-   or a salt thereof, wherein:    -   R₁′ is selected from H or optionally substituted C₁-C₃ straight        or branched alkyl;    -   R²¹ is selected from —NR₁′—C(O)—, —NR₁′—S(O)₂—,        —NR₁′—C(O)—NR₁′—, —NR₁′—C(S)—NR₁′—, —NR₁′—C(S)—NR₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—NR₁′—, —NR₁′—C(═NR₁′)—NR₁′—, —C(O)—NR₁′—,        —C(O)—NR₁′—S(O)₂—, —NR₁′—, —CR₁′R₁′—, —NR₁′—C(O)—CR₁′═CR₁′—,        —NR₁′—S(O)₂—NR₁′—, —NR₁′—C(O)—NR₁′—S(O)₂—,        —NR₁′—CR₁′R₁′—C(O)—NR₁′—, —CR₁′R₁′—C(O)—NR₁′—,        —NR₁′—C(O)—CR₁′═CR₁′—CR₁′R₁′—, —NR₁′—C(═N—CN)—NR₁′—,        —NR₁′—C(O)—CR₁′R₁′—O—, —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—O—,        —NR₁′—S(O)₂—CR₁′R₁′—, —NR₁′—S(O)₂—CR₁′R₁′—CR₁′R₁′—, or        —NR₁′—C(O)—CR₁′R₁′—; and    -   R³¹ is selected from an optionally substituted monocyclic or        bicyclic aryl, or an optionally substituted monocyclic or        bicyclic heteroaryl, with the provisos that:    -   when R²¹ is —NH—C(O)—, R³¹ is not unsubstituted furyl,        5-bromofuryl, unsubstituted phenyl, phenyl monosubstituted with        halo or methyl, 3- or 4-methoxyphenyl, 4-butoxyphenyl,        4-t-butylphenyl, 3-trifluoromethylphenyl, 2-benzoylphenyl, 2- or        4-ethoxyphenyl, 2,3-, 2,4-, 3,4-, or 3,5-dimethoxyphenyl,        3,4,5-trimethoxyphenyl, 2,4- or 2-6 difluorophenyl,        3,4-dioxymethylene phenyl, 3,4- or 3,5-dimethlyphenyl,        2-chloro-5-bromophenyl, 2-methoxy-5-chlorophenyl, unsubstituted        quinolinyl, thiazolyl substituted simultaneously with methyl and        phenyl, or ethoxy-substituted pyridinyl;    -   when R²¹ is —NH—C(O)—CH(CH₂—CH₃)—, R³¹ is not unsubstituted        phenyl;    -   when R²¹ is —NH—C(O)—CH₂—, R³¹ is not unsubstituted phenyl,        3-methylphenyl, 4-chlorophenyl, 4-ethoxyphenyl, 4-fluorophenyl        or 4-methoxyphenyl;    -   when R²¹ is —NH—C(O)—CH₂—O—, R³¹ is not unsubstituted phenyl or        4-chlorophenyl; and    -   when R²¹ is —NH—S(O)₂—, R³¹ is not 3,4-dioxymethylene phenyl,        2,4,5-trimethylphenyl, 2,4,6-trimethylphenyl, 2,4- or        3,4-dimethylphenyl, 2,5-difluorophenyl, 2,5- or        3,4-dimethoxyphenyl, fluorophenyl, 4-chlorophenyl,        4-bromophenyl, 4-ethylphenyl, 4-methylphenyl,        3-methyl-4-methoxyphenyl, unsubstituted phenyl, unsubstituted        pyridinyl, unsubstituted thienyl, chloro-substituted thienyl, or        methyl-substituted benzothiazolyl.

In certain embodiments, R₁′ is selected from H or optionally substitutedC₁-C₃ straight or branched alkyl;

-   -   R²¹ is selected from —NR₁′—C(O)—, —NR₁′—S(O)₂—,        —NR₁′—C(O)—NR₁′—, —NR₁′—C(S)—NR₁′—, —NR₁′—C(S)—NR₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—NR₁′—, —NR₁′—C(═NR₁′)—NR₁′—, —C(O)—NR₁′—,        —C(O)—NR₁′—S(O)₂—, —NR₁′—, —CR₁′R₁′—, —NR₁′—C(O)—CR₁′═CR₁′—,        —NR₁′—S(O)₂—NR₁′—, —NR₁′—C(O)—NR₁′—S(O)₂—,        —NR₁′—CR₁′R₁′—C(O)—NR₁′—, —CR₁′R₁′—C(O)—NR₁′—,        —NR₁′—C(O)—CR₁′═CR₁′—CR₁′R₁′—, —NR₁′—C(═N—CN)—NR₁′—,        —NR₁′—C(O)—CR₁′R₁′—O—, —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—O—,        —NR₁′—S(O)₂—CR₁′R₁′—, —NR₁′—S(O)₂—CR₁′R₁′—CR₁′R₁′—, or        —NR₁′—C(O)—CR₁′R₁′—; and    -   R³¹ is selected from a monocyclic or bicyclic aryl or a        monocyclic or bicyclic heteroaryl, and comprises a solubilizing        group substituent.

In certain embodiments, R³¹ is selected from phenyl, naphthyl,pyrazolyl, furyl, thienyl, pyridyl, isoxazolyl, benzopyrazolyl,benzofuryl, benzothienyl, quinolinyl, benzoisoxazolyl, or

and R³¹ is optionally substituted.

In certain embodiments, R²¹ is selected from —NH—C(O)—,NH—C(O)—CH₂—CH(CH₃)—O, —NH—C(O)—NH—, —NH—C(S)—NH—, —NH—C(S)—NH—CH₂—, or—NH—S(O)₂—; and

-   -   R³¹ is selected from an optionally substituted phenyl, an        optionally substituted naphthyl, or an optionally substituted        heteroaryl.

In certain such embodiments, particularly when R²¹ is —NH—C(O)—, R³¹ isselected from R³¹ is selected from unsubstituted phenyl,3-methoxyphenyl, 4-methoxyphenyl, 2,3 dimethoxyphenyl,2,4-dimethoxyphenyl, 2,5-bis(trifluoromethyl)phenyl,3,4-dimethoxyphenyl, 3,5-dimethoxyphenyl, 3,4,5-trimethoxyphenyl,2,3,4-trimethoxyphenyl, 2-methoxy-4-methylphenyl, 2-phenoxyphenyl,3-dimethylaminophenyl, 4-dimethylaminophenyl, unsubstituted 2-furanyl,unsubstituted 2-thienyl,

In one aspect, the invention provides sirtuin-modulating compounds ofStructural Formula (XIV):

-   or a salt thereof, wherein:    -   each of R²³ and R²⁴ is independently selected from H, —CH₃ or a        solubilizing group;    -   R²⁵ is selected from H or a solubilizing group; and    -   R¹⁹ is selected from:

-   -   wherein:        -   each Z₁₀, Z₁₁, Z₁₂ and Z₁₃ is independently selected from N,            CR²⁰, or CR₁′; and        -   each Z₁₄, Z₁₅ and Z₁₆ is independently selected from N,            NR₁′, S, O, CR²⁰, or CR₁′, wherein:        -   zero to two of Z₁₀, Z₁₁, Z₁₂ or Z₁₃ are N;        -   at least one of Z₁₄, Z₁₅ and Z₁₆ is N, NR₁′, O or S;        -   zero to one of Z₁₄, Z₁₅ and Z₁₆ is S or O;        -   zero to two of Z₁₄, Z₁₅ and Z₁₆ are N or NR₁′;        -   zero to one R²⁰ is a solubilizing group; and        -   zero to one R₁′ is an optionally substituted C₁-C₃ straight            or branched alkyl;        -   each R²⁰ is independently selected from H or a solubilizing            group;    -   R²¹ is selected from —NR₁′—C(O)—, —NR₁′—S(O)₂—,        —NR₁′—C(O)—NR₁′—, —NR₁′—C(S)—NR₁′—, —NR₁′—C(S)—NR₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—NR₁′—, —NR₁′—C(═NR₁′)—NR₁′—, —C(O)—NR₁′—,        —C(O)—NR₁′—S(O)₂—, —NR₁′—, —CR₁′R₁′—, —NR₁′—C(O)—CR₁′—CR₁′—,        —NR₁′—S(O)₂—NR₁′—, —NR₁′—C(O)—NR₁′—S(O)₂—,        —NR₁′—CR₁′R₁′—C(O)—NR₁′—, —CR₁′R₁′—C(O)—NR₁′—,        —NR₁′—C(O)—CR₁′═CR₁′—CR₁′R₁′—, —NR₁′—C(═N—CN)—NR₁′—,        —NR₁′—C(O)—CR₁′R₁′—O—, —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—O—,        —NR₁′—S(O)₂—CR₁′R₁′—, —NR₁′—S(O)₂—CR₁′R₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—, —NR₁′—C(S)—NR₁′—CR₁′R₁′—CR₁′R₁′—,        —NR₁′—C(O)—O— or —NR₁′—C(O)—CR₁′R₁′—; and    -   each R₁′ is independently selected from H or optionally        substituted C₁-C₃ straight or branched alkyl; and    -   R³¹ is selected from an optionally substituted monocyclic or        bicyclic aryl, or an optionally substituted monocyclic or        bicyclic heteroaryl,        -   wherein when R¹⁹ is

-   -   -    R²¹ is —NH—C(O)— and R²⁵ is —H, R³¹ is not an optionally            substituted phenyl group, and wherein said compound is not            2-chloro-N-[3-[3-(cyclohexylamino)imidazo[1,2-a]pyridin-2-yl]phenyl]-4-nitrobenzamide.

In certain embodiments, each of R²³ and R²⁴ is independently selectedfrom H, —CH₃ or a solubilizing group;

-   -   R²⁵ is selected from H, or a solubilizing group; and    -   R¹⁹ is selected from:

-   -   wherein:        -   each Z₁₀, Z₁₁, Z₁₂ and Z₁₃ is independently selected from N,            CR²⁰, or CR₁′; and        -   each Z₁₄, Z₁₅ and Z₁₆ is independently selected from N,            NR₁′, S, O, CR²⁰, or CR₁′,    -   wherein:    -   zero to two of Z₁₀, Z₁₁, Z₁₂ or Z₁₃ are N;    -   at least one of Z₁₄, Z₁₅ and Z₁₆ is N, NR₁′, O or S;    -   zero to one of Z₁₄, Z₁₅ and Z₁₆ is S or O;    -   zero to two of Z₁₄, Z₁₅ and Z₁₆ are N or NR₁′;    -   zero to one R²⁰ is a solubilizing group; and    -   zero to one R₁′ is an optionally substituted C₁-C₃ straight or        branched alkyl;    -   each R²⁰ is independently selected from H or a solubilizing        group;    -   R²¹ is selected from —NR₁′—C(O)—, —NR₁′—S(O)₂—,        —NR₁′—C(O)—NR₁′—, —NR₁′—C(S)—NR₁′—, —NR₁′—C(S)—NR₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—NR₁′—, —NR₁′—C(═NR₁′)—NR₁′—, —C(O)—NR₁′—,        —C(O)—NR₁′—S(O)₂—, —NR₁′—, —CR₁′R₁′—, —NR₁′—C(O)—CR₁′═CR₁′—,        —NR₁′—S(O)₂—NR₁′—, —NR₁′—C(O)—NR₁′—S(O)₂—,        —NR₁′—CR₁′R₁′—C(O)—NR₁′—, —CR₁′R₁′—C(O)—NR₁′—,        —NR₁′—C(O)—CR₁′═CR₁′—CR₁′R₁′—, —NR₁′—C(═N—CN)—NR₁′—,        —NR₁′—C(O)—CR₁′R₁′—O—, —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—O—,        —NR₁′—S(O)₂—CR₁′R₁′—, —NR₁′—S(O)₂—CR₁′R₁′— CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—NR₁′—C(S)—NR₁′—CR₁′R₁′—CR₁′R₁′—,        —NR₁′—C(O)—O— or —NR₁′—C(O)—CR₁′R₁′— (particularly —NH—C(O)—);        and    -   each R₁′ is independently selected from H or optionally        substituted C₁-C₃ straight or branched alkyl; and    -   R³¹ is selected from an optionally substituted monocyclic or        bicyclic aryl, or an optionally substituted monocyclic or        bicyclic heteroaryl.

In certain such embodiments, R³¹ is not 2,4-dimethoxyphenyl.

Typically, R²⁵ is selected from H, —CH₂—N(CH₃)₂, or

Typically, R²³ and R²⁴ are H.

Typically, R¹⁹ is selected from phenyl, pyridyl, thienyl or furyl,particularly optionally substituted phenyl. Preferably, a phenyl isoptionally substituted with:

-   -   a) up to three —O—CH₃ groups; or    -   b) one —N(CH₃)₂ group.

In certain embodiments, each of R²³ and R²⁴ is independently selectedfrom H, —CH₃ or a solubilizing group;

-   -   R²⁵ is selected from H, or a solubilizing group; and    -   R¹⁹ is selected from:

-   -   wherein:        -   each Z₁₀, Z₁₀, Z₁₂ and Z₁₃ is independently selected from N,            CR²⁰, or CR₁′; and        -   each Z₁₄, Z₁₅ and Z₁₆ is independently selected from N,            NR₁′, S, O, CR²⁰, or CR₁′, wherein:        -   zero to two of Z₁₀, Z₁₁, Z₁₂ or Z₁₃ are N;        -   at least one of Z₁₄, Z₁₅ and Z₁₆ is N, NR₁′, O or S;        -   zero to one of Z₁₄, Z₁₅ and Z₁₆ is S or O;        -   zero to two of Z₁₄, Z₁₅ and Z₁₆ is N or NR₁′;        -   zero to one R²⁰ is a solubilizing group; and        -   zero to one R₁′ is an optionally substituted C₁-C₃ straight            or branched alkyl;        -   each R²⁰ is independently selected from H or a solubilizing            group;    -   R²¹ is selected from —NR₁′—C(O)—, —NR₁′—S(O)₂—,        —NR₁′—C(O)—NR₁′—, —NR₁′—C(S)—NR₁′—, —NR₁′—C(S)—NR₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—NR₁′—, —NR₁′—C(═NR₁′)—NR₁′—, —C(O)—NR₁′—,        —C(O)—NR₁′—S(O)₂—, —NR₁′—, —CR₁′R₁′—, —NR₁′—C(O)—CR₁′═CR₁′—,        —NR₁′—S(O)₂—NR₁₁′—, —NR₁′—C(O)—NR₁′—S(O)₂—,        —NR₁′—CR₁′R₁′—C(O)—NR₁′—, —CR₁′R₁′—C(O)—NR₁′—,        —NR₁′—C(O)—CR₁′═CR₁′—CR₁′R₁′—, —NR₁′—C(═N—CN)—NR₁′—,        —NR₁′—C(O)—CR₁′R₁′—O—, —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—O—,        —NR₁′—S(O)₂—CR₁′R₁′—, —NR₁′—S(O)₂—CR₁′R₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—, —NR₁′—C(S)—NR₁′—CR₁′R₁′—CR₁′R₁′—,        —NR₁′—C(O)—O— or —NR₁′—C(O)—CR₁′R₁′— (particularly —NH—C(O)—);        and    -   each R₁′ is independently selected from H or optionally        substituted C₁-C₃ straight or branched alkyl; and    -   R³¹ is selected from an optionally substituted monocyclic or        bicyclic aryl, or an optionally substituted monocyclic or        bicyclic heteroaryl, wherein when R¹⁹ is phenyl, at least one of        R²³, R²⁴, or R²⁵ is a solubilizing group and wherein said        compound is not        2-chloro-N-[3-[3-(cyclohexylamino)imidazo[1,2-a]pyridin-2-yl]phenyl]-4-nitrobenzamide.

Typically, R²⁵ is selected from H, —CH₂—N(CH₃)₂, or

Typically, R²³ and R²⁴ are H.

Typically, R¹⁹ is selected from phenyl, pyridyl, thienyl or furyl,particularly optionally substituted phenyl. Preferably, a phenyl isoptionally substituted with:

-   -   b) up to three —O—CH₃ groups; or    -   b) one —N(CH₃)₂ group.

In another aspect, the invention provides sirtuin-modulating compoundsof Structural Formula (XV):

-   or a salt thereof, wherein:    -   R²¹ is selected from —NR₁′—C(O)—, —NR₁′—S(O)₂—,        —NR₁′—C(O)—NR₁′—, —NR₁′—C(S)—NR₁′—, —NR₁′—C(S)—NR₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—NR₁′—, —NR₁′—C(═NR₁′)—NR₁′—, —C(O)—NR₁′—,        —C(O)—NR₁′—S(O)₂—, —NR₁′—, —CR₁′R₁′—, —NR₁′—C(O)—CR₁′═CR₁′—,        —NR₁′—S(O)₂—NR₁′—, —NR₁′—C(O)—NR₁′—S(O)₂—,        —NR₁′—CR₁′R₁′—C(O)—NR₁′—, —CR₁′R₁′—C(O)—NR₁′—,        —NR₁′—C(O)—CR₁′═CR₁′—CR₁′R₁′—, —NR₁′—C(═N—CN)—NR₁′—,        —NR₁′—C(O)—CR₁′R₁′—O—, —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—O—,        —NR₁′—S(O)₂—CR₁′R₁′—, —NR₁′—S(O)₂—CR₁′R₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—, —NR₁′—C(S)—NR₁′—CR₁′R₁′—CR₁′R₁′—,        —NR₁′—C(O)—O— or —NR₁′—C(O)—CR₁′R₁′— (particularly —NH—C(O)—);        and    -   each R₁′ is independently selected from H or optionally        substituted C₁-C₃ straight or branched alkyl; and    -   R³² is selected from an optionally substituted bicyclic aryl, or        an optionally substituted monocyclic or bicyclic heteroaryl,        wherein:    -   when R²¹ is —NH—C(O)—, R³² is not unsubstituted 2-furyl,        2-(3-bromofuryl), unsubstituted 2-thienyl, unsubstituted        3-pyridyl, unsubstituted 4-pyridyl,

-   -   when R²¹ is —NR₁′—S(O)₂—, R³² is not unsubstituted 2-thienyl or        unsubstituted naphthyl.

In yet another aspect, the invention provides sirtuin-modulatingcompounds of Structural Formula (XVI):

-   or a salt thereof, wherein:    -   R²¹ is selected from —NR₁′—C(O)—, —NR₁′—S(O)₂—,        —NR₁′—C(O)—NR₁′—, —NR₁′—C(S)—NR₁′—, —NR₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—NR₁′—, —NR₁′—C(═NR₁′)—NR₁′—, —C(O)—NR₁′—,        —C(O)—NR₁′—S(O)₂—, —NR₁′—, —CR₁′R₁′—, —NR₁′—C(O)—CR₁′═CR₁′—,        —NR₁′—S(O)₂—NR₁′—, —NR₁′—C(O)—NR₁′—S(O)₂—,        —NR₁′—CR₁′R₁′—C(O)—NR₁′—, —CR₁′R₁′—C(O)—NR₁′—,        —NR₁′—C(O)—CR₁′R₁′—O—, —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—O—,        —NR₁′—S(O)₂—CR₁′R₁′—, —NR₁′—S(O)₂—CR₁′R₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′═CR₁′—CR₁′R₁′—, —NR₁′—C(═N—CN)—NR₁′—,        —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—, —NR₁′—C(S)—NR₁′—CR₁′R₁′—CR₁′R₁′—,        —NR₁′—C(O)—O— or —NR₁′—C(O)—CR₁′R₁′— (particularly —NH—C(O)—);        and    -   each R₁′ is independently selected from H or optionally        substituted C₁-C₃ straight or branched alkyl; and    -   R³³ is an optionally substituted phenyl, wherein:    -   when R²¹ is —NH—C(O)—, R³³ is a substituted phenyl other than        phenyl singly substituted with halo, methyl, nitro or methoxy;        2-carboxyphenyl; 4-n-pentylphenyl; 4-ethoxyphenyl;        2-carboxy-3-nitrophenyl; 2-chloro-4-nitrophenyl;        2-methoxy-5-ethylphenyl; 2,4-dimethoxyphenyl;        3,4,5-trimethoxyphenyl; 2,4 dichlorophenyl; 2,6-difluorophenyl;        3,5-dinitrophenyl; or 3,4-dimethylphenyl;    -   when R²¹ is —NR₁′—C(O)—CR₁′R₁′— or —NH—C(O)—CH(CH₃)—O, R³³ is a        substituted phenyl;    -   when R²¹ is —NH—C(O)—CH₂, R³³ is not unsubstituted phenyl,        4-methoxyphenyl; 3,4-dimethoxyphenyl or 4-chlorophenyl;    -   when R²¹ is —NH—C(O)—CH₂—O, R³³ is not        2,4-bis(1,1-dimethylpropyl)phenyl;    -   when R²¹ is —NH—C(O)—NH—, R³³ is not 4-methoxyphenyl; and    -   when R²¹ is —NH—S(O)₂—, R³³ is a substituted phenyl other than        3-methylphenyl, 3-trifluoromethylphenyl, 2,4,5- or        2,4,6-trimethylphenyl, 2,4- or 3,4-dimethylphenyl, 2,5- or        3,4-dimethoxyphenyl, 2,5-dimethoxy-4-chlorophenyl,        3,6-dimethoxy, 4-methylphenyl, 2,5- or 3,4-dichlorophenyl,        2,5-diethoxyphenyl, 2-methyl -5-nitrophenyl,        2-ethoxy-5-bromophenyl, 2-methoxy-5-bromophenyl,        2-methoxy-3,4-dichlorophenyl, 2-methoxy-4-methyl-5-bromophenyl,        3,5-dinitro-4-methylphenyl, 3-methyl-4-methoxyphenyl,        3-nitro-4-methylphenyl, 3-methoxy-4-halophenyl,        3-methoxy-5-chlorophenyl, 4-n-butoxyphenyl, 4-halophenyl,        4-ethylphenyl, 4-methylphenyl, 4-nitrophenyl, 4-ethoxyphenyl,        4-acetylaminophenyl, 4-methoxyphenyl, 4-t -butylphenyl, or        para-biphenyl.

In certain embodiments, R²¹ is selected from —NR²²—C(O)—, —NR₁′—S(O)₂—,—NR₁′—C(O)—NR₁′—, —NR₁′—C(S)—NR₁′—, —NR₁′—C(S)—NR₁′—CR₁′R₁′—,—NR₁′—C(O)—CR₁′R₁′—NR₁′—, —NR₁′—C(═NR₁′)—NR₁′—, —C(O)—NR₁′—,—C(O)—NR₁′—S(O)₂—, —NR₁′—, —CR₁′R₁′—, —NR₁′—C(O)—CR₁′═CR₁′—,—NR₁′—S(O)₂—NR₁′—, —NR₁′—C(O)—NR₁′—S(O)₂—, —NR₁′—CR₁′R₁′—C(O)—NR₁′—,—CR₁′R₁′—C(O)—NR₁′—, —NR₁′—C(O)—CR₁′R₁′—O—,—NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—O—, —NR₁′—S(O)₂—CR₁′R₁′—,—NR₁′—S(O)₂—CR₁′R₁′—CR₁′R₁′—, —NR₁′—C(O)—CR₁′—CR₁′—CR₁′R₁′—,—NR₁′—C(═N—CN)—NR₁′—, or —NR₁′—C(O)—CR₁′R₁′—; and

-   -   each R₁′ is independently selected from H or optionally        substituted C₁-C₃ straight or branched alkyl;    -   R²² is an optionally substituted C₁-C₃ straight or branched        alkyl; and    -   R³³ is phenyl comprising a solubilizing group substituent,        wherein: when R²¹ is —NH—S(O)₂ said phenyl comprises an        additional substituent.

In certain embodiments, R²¹ is selected from —NR²²—C(O)—,—NR₁′—C(S)—NR₁′—, —NR₁′—C(S)—NR₁′—CR₁′R₁′—, —NR₁′—C(O)—CR₁′R₁′—NR₁′—,—NR₁′—C(═NR₁′)—NR₁′—, —C(O)—NR₁′—, —C(O)—NR₁′—S(O)₂—, —NR₁′—, —CR₁′R₁′—,—NR₁′—C(O)—CR₁′═CR₁′—, —NR₁′—S(O)₂—NR₁′—, —NR₁′—C(O)—NR₁′—S(O)₂—,—NR₁′—CR₁′R₁′—C(O)—NR₁′—, —CR₁′R₁′—C(O)—NR₁′—,—NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—O—, —NR₁′—S(O)₂—CR₁′R₁′—,—NR₁′—S(O)₂—CR₁′R₁′—CR₁′R₁′—, —NR₁′—C(O)—CR₁′═CR₁′—CR₁′R₁′—, or—NR₁′—C(═N—CN)—NR₁′—,

-   -   each R₁′ is independently selected from H or optionally        substituted C₁-C₃ straight or branched alkyl; and    -   R²² is an optionally substituted C₁-C₃ straight or branched        alkyl.

In certain embodiments, R³³ is optionally substituted on up to threecarbon atoms with a substituent independently selected from —O—CH₃,—CH₃, —N(CH₃)₂, —S(CH₃), or CN; or substituted on adjacent carbon atomswith

bridging said adjacent carbon atoms.

In a further embodiment, the invention provides sirtuin-modulatingcompounds of Structural Formula (XVII):

-   or a salt thereof, wherein:    -   each of R²³ and R²⁴ is independently selected from H or —CH₃,        wherein at least one of R²³ and R²⁴ is H; and    -   R²⁹ is phenyl substituted with:    -   a) two —O—CH₃ groups;    -   b) three —O—CH₃ groups located at the 2, 3 and 4 positions; or    -   c) one —N(CH₃)₂ group; and;    -   d) when R²³ is CH₃, one —O—CH₃ group at the 2 or 3 position,    -   wherein R²⁹ is optionally additionally substituted with a        solubilizing group.

In certain embodiments, R²⁹ is phenyl substituted with:

-   -   a) three —O—CH₃ groups located at the 2, 3 and 4 positions; or    -   b) one —N(CH₃)₂ group.

In one aspect, the invention provides sirtuin-modulating compounds ofStructural Formula (XVIII):

-   or a salt thereof, wherein    -   R¹⁹ is selected from:

-   -   wherein:        -   each Z₁₀, Z₁₁, Z₁₂ and Z₁₃ is independently selected from N,            CR²⁰, or CR₁′; and        -   each Z₁₄, Z₁₅ and Z₁₆ is independently selected from N,            NR₁′, S, O, CR²⁰, or CR₁′,    -   wherein:    -   zero to two of Z₁₀, Z₁₁, Z₁₂ or Z₁₃ are N;    -   at least one of Z₁₄, Z₁₅ and Z₁₆ is N, NR₁′, S or O;    -   zero to one of Z₁₄, Z₁₅ and Z₁₆ is S or O;    -   zero to two of Z₁₄, Z₁₅ and Z₁₆ are N or NR₁′;    -   zero to one R²⁰ is a solubilizing group; and    -   zero to one R₁′ is an optionally substituted C₁-C₃ straight or        branched alkyl;    -   each R²⁰ is independently selected from H or a solubilizing        group;    -   R²¹ is selected from —NR₁′—C(O)—, —NR₁′—S(O)₂—,        —NR₁′—C(O)—NR₁′—, —NR₁′—C(S)—NR₁′—, —NR₁′—C(S)—NR₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—NR₁′—, —NR₁′—C(═NR₁′)—NR₁′—, —C(O)—NR₁′—,        —C(O)—NR₁′—S(O)₂—, —NR₁′—, —CR₁′R₁′—, —NR₁′—C(O)—CR₁′═CR₁′—,        —NR₁′—S(O)₂—NR₁′—, —NR₁′—C(O)—NR₁′—S(O)₂—,        —NR₁′—CR₁′R₁′—C(O)—NR₁′—, —CR₁′R₁′—C(O)—NR₁′—,        —NR₁′—C(O)—CR₁′═CR₁′—CR₁′R₁′—, —NR₁′—C(═N—CN)—NR₁′—,        —NR₁′—C(O)—CR₁′R₁′—O—, —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—O—,        —NR₁′—S(O)₂—CR₁′R₁′—, —NR₁′—S(O)₂—CR₁′R₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—; —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—,        —NR₁′—C(S)—NR₁′—CR₁′R₁′—CR₁′R₁′—, —NR₁′—C(O)—O—,

-   -    wherein each R₁′ is independently selected from H or optionally        substituted C₁-C₃ straight or branched alkyl; and    -   R³¹ is selected from an optionally substituted monocyclic or        bicyclic aryl, or an optionally substituted monocyclic or        bicyclic heteroaryl, with the proviso that when R¹⁹ is

-   -    Z₁₀, Z₁₁, Z₁₂ and Z₁₃ are each CH, R²⁰ is H, and R²¹ is        —NHC(O)—, R³¹ is not an optionally substituted phenyl.

In certain embodiments, R¹⁹ is selected from:

-   -   wherein:        -   each Z₁₀, Z₁₀, Z₁₂ and Z₁₃ is independently selected from N,            CR²⁰, or CR₁′; and        -   each Z₁₄, Z₁₅ and Z₁₆ is independently selected from N,            NR₁′, S, O, CR²⁰, or CR₁′,    -   wherein:    -   zero to two of Z₁₀, Z₁₁, Z₁₂ or Z₁₃ are N;    -   at least one of Z₁₄, Z₁₅ and Z₁₆ is N, NR₁′, O or S;    -   zero to one of Z₁₄, Z₁₅ and Z₁₆ is S or O;    -   zero to two of Z₁₄, Z₁₅ and Z₁₆ are N or NR₁′;    -   zero to one R²⁰ is a solubilizing group; and    -   zero to one R₁′ is an optionally substituted C₁-C₃ straight or        branched alkyl;    -   each R²⁰ is independently selected from H or a solubilizing        group;    -   R²¹ is selected from —NR₁′—C(O)—, —NR₁′—S(O)₂—,        —NR₁′—C(O)—NR₁′—, —NR₁′—C(S)—NR₁′—, —NR₁′—C(S)—NR₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—NR₁′—, —NR₁′—C(═NR₁′)—NR₁′—, —C(O)—NR₁′—,        —C(O)—NR₁′—S(O)₂—, —NR₁′—, —CR₁′R₁′—, —NR₁′—C(O)—CR₁′═CR₁′—,        —NR₁′—S(O)₂—NR₁′—, —NR₁′—C(O)—NR₁′—S(O)₂—,        —NR₁′—CR₁′R₁′—C(O)—NR₁′—, —CR₁′R₁′—C(O)—NR₁′—,        —NR₁′—C(O)—CR₁′═CR₁′—CR₁′R₁′—, —NR₁′—C(═N—CN)—NR₁′—,        —NR₁′—C(O)—CR₁′R₁′—O—, —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—O—,        —NR₁′—S(O)₂—CR₁′R₁′—, —NR₁′—S(O)₂—CR₁′R₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—; —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—,        —NR₁′—C(S)—NR₁′—CR₁′R₁′—CR₁′R₁′—, —NR₁′—C(O)—O—,

-   -   each R₁′ is independently selected from H or optionally        substituted C₁-C₃ straight or branched alkyl; and    -   R³¹ is selected from an optionally substituted monocyclic or        bicyclic aryl, or an optionally substituted monocyclic or        bicyclic heteroaryl.

In certain such embodiments, compounds of Structural Formula (XVIII)have the formula:

-   or a salt thereof, wherein    -   R²⁰ is selected from H or a solubilizing group;    -   R²¹ is selected from —NH—C(O)—, or —NH—C(O)—CH₂—; and    -   R³¹ is selected from an optionally substituted monocyclic or        bicyclic aryl, or an optionally substituted monocyclic or        bicyclic heteroaryl.

Typically, R¹⁹ in compounds of Structural Formula (XVIII) is selectedfrom phenyl, pyridyl, thienyl or furyl, particularly optionallysubstituted phenyl.

Typically, R²⁰ is selected from H, —CH₂—N(CH₃)₂,

Typically, R³¹ is selected from phenyl, pyrazolyl, furyl, pyridyl,pyrimidinyl, thienyl, naphthyl, benzopyrazolyl, benzofuryl, quinolinyl,quinoxalinyl, or benzothienyl and wherein R³¹ is optionally substituted.

Typically, R²¹ is selected from —NH—C(O)— or —NH—C(O)—CH₂—.

In certain such embodiments, when R²¹ is —NR₁′—C(O)—, R³¹ is not4-cyanophenyl or

and/or when R²¹ is —NR₁′—S(O)₂—, R³¹ is not 4-methoxyphenyl or4-t-butylphenyl.

In certain such embodiments, when R¹⁹ is

and R²¹ is —NR₁′—C(O)—, R³¹ is not 4-cyanophenyl or

and/or when R¹⁹ is

and R²¹ is —NR₁′—S(O)₂—, R³¹ is not 4-methoxyphenyl or 4-t-butylphenyl.

In another aspect, the invention provides sirtuin-modulating compoundsof Structural Formula (XX):

-   or a salt thereof, wherein    -   R¹⁹ is selected from:

-   -   wherein:        -   each Z₁₀, Z₁₁, Z₁₂ and Z₁₃ is independently selected from N,            CR²⁰, or CR₁′; and        -   each Z₁₄, Z₁₅ and Z₁₆ is independently selected from N,            NR₁′, S, O, CR²⁰, or CR₁′,    -   wherein:        -   zero to two of Z₁₀, Z₁₁, Z₁₂ or Z₁₃ are N;        -   at least one of Z₁₄, Z₁₅ and Z₁₆ is N, NR₁′, O or S;        -   zero to one of Z₁₄, Z₁₅ and Z₁₆ is S or O;        -   zero to two of Z₁₄, Z₁₅ and Z₁₆ are N or NR₁′;        -   zero to one R²⁰ is a solubilizing group; and        -   zero to one R₁′ is an optionally substituted C₁-C₃ straight            or branched alkyl;    -   each R²⁰ is independently selected from H or a solubilizing        group;    -   R^(20a) is independently selected from H or a solubilizing        group;    -   R²¹ is selected from —NR₁′—C(O)—, —NR₁′—S(O)₂—,        —NR₁′—C(O)—NR₁′—, —NR₁′—C(S)—NR₁′—, —NR₁′—C(S)—NR₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—NR₁′—, —NR₁′—C(═NR₁′)—NR₁′—, —C(O)—NR₁′—,        —C(O)—NR₁′—S(O)₂—, —NR₁′—, —CR₁′R₁′—, —NR₁′—C(O)—CR₁′═CR₁′—,        —NR₁′—S(O)₂—NR₁′—, —NR₁′—C(O)—NR₁′—S(O)₂—,        —NR₁′—CR₁′R₁′—C(O)—NR₁′—, —CR₁′R₁′—C(O)—NR₁′—,        —NR₁′—C(O)—CR₁′═CR₁′—CR₁′R₁′—, —NR₁′—C(═N—CN)—NR₁′—,        —NR₁′—C(O)—CR₁′R₁′—O—, —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—O—,        —NR₁′—S(O)₂—CR₁′R₁′—, —NR₁′—S(O)₂—CR₁′R₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—; —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—,        —NR₁′—C(S)—NR₁′—CR₁′R₁′—CR₁′R₁′—, —NR₁′—C(O)—O—,

-   -   wherein        -   each R₁′ is independently selected from H or optionally            substituted C₁-C₃ straight or branched alkyl; and    -   R³¹ is selected from an optionally substituted monocyclic or        bicyclic aryl, or an optionally substituted monocyclic or        bicyclic heteroaryl, wherein when R¹⁹ is

-   -    and Z₁₀, Z₁₁, Z₁₂ and Z₁₃ are each CH, R^(20a) is a        solubilizing group.

Typically, R¹⁹ in compounds of Structural Formula (XX) is selected fromphenyl, pyridyl, thienyl or furyl, particularly optionally substitutedphenyl.

Typically, R^(20a) is selected from H, —CH₂—N(CH₃)₂,

Typically, R³¹ is selected from phenyl, pyrazolyl, furyl, pyridyl,pyrimidinyl, thienyl, naphthyl, benzopyrazolyl, benzofuryl, quinolinyl,quinoxalinyl, or benzothienyl and wherein R³¹ is optionally substituted.

Typically, R²¹ is selected from —NH—C(O)— or —NH—C(O)—CH₂—.

In yet another aspect, the invention provides sirtuin-modulatingcompounds of Structural Formula (XXI):

-   or a salt thereof, wherein    -   R²¹ is selected from —NR₁′—C(O)—, —NR₁′—S(O)₂—,        —NR₁′—C(O)—NR₁′—, —NR₁′—C(S)—NR₁′—, —NR₁′—C(S)—NR₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—NR₁′—, —NR₁′—C(═NR₁′)—NR₁′—, —C(O)—NR₁′—,        —C(O)—NR₁′—S(O)₂—, —NR₁′—, —CR₁′R₁′—, —NR₁′—C(O)—CR₁′═CR₁′—,        —NR₁′—S(O)₂—NR₁′—, —NR₁′—C(O)—NR₁′—S(O)₂—,        —NR₁′—CR₁′R₁′—C(O)—NR₁′—, —CR₁′R₁′—C(O)—NR₁′—,        —NR₁′—C(O)—CR₁′═CR₁′—CR₁′R₁′—, —NR₁′—C(═N—CN)—NR₁′—,        —NR₁′—C(O)—CR₁′R₁′—O—, —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—O—,        —NR₁′—S(O)₂—CR₁′R₁′—, —NR₁′—S(O)₂—CR₁′R₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—; —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—,        —NR₁′—C(S)—NR₁′—CR₁′R₁′—CR₁′R₁′—, —NR₁′—C(O)—O—,

-   -   wherein        -   each R₁′ is independently selected from H or optionally            substituted C₁-C₃ straight or branched alkyl; and    -   R³² is an optionally substituted monocyclic or bicyclic        heteroaryl, or an optionally substituted bicyclic aryl, wherein:    -   when R²¹ is —NH—C(O)—CH₂—, R³² is not unsubstituted thien-2-yl;    -   when R²¹ is —NH—C(O)—, R³² is not furan-2-yl, 5-bromofuran-2-yl,        or 2-phenyl-4-methylthiazol-5-yl;    -   when R²¹ is —NH—S(O)₂—, R³² is not unsubstituted naphthyl or        5-chlorothien-2-yl.

In certain embodiments, R³² is selected from pyrrolyl, pyrazolyl,pyrazinyl, furyl, pyridyl, pyrimidinyl, or thienyl, and R³² isoptionally substituted and is optionally benzofused.

In certain embodiments, R²¹ is selected from —NR₁′—C(O)—, —NR₁′—S(O)₂—,—NR₁′—C(O)—NR₁′—, —NR₁′—C(S)—NR₁′—, —NR₁′—C(S)—NR₁′—CR₁′R₁′—,—NR₁′—C(O)—CR₁′R₁′—NR₁′—, —NR₁′—C(═NR₁′)—NR₁′—, —C(O)—NR₁′—,—C(O)—NR₁′—S(O)₂, —NR₁′—, —CR₁′R₁′—, —NR₁′—C(O)—CR₁′═CR₁′—,—NR₁′—S(O)₂—NR₁′—, —NR₁′—C(O)—NR₁′—S(O)₂—, —NR₁′—CR₁′R₁′—C(O)—NR₁′—,—CR₁′R₁′—C(O)—NR₁′—, —NR₁′—C(O)—CR₁′═CR₁′—CR₁′R₁′—,—NR₁′—C(═N—CN)—NR₁′—, —NR₁′—C(O)—CR₁′R₁′—O—,—NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—O—, —NR₁′—S(O)₂—CR₁′R₁′—,—NR₁′—S(O)₂—CR₁′R₁′—CR₁′R₁′—, —NR₁′—C(O)—CR₁′R₁′—;—NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—, —NR₁′—C(S)—NR₁′—CR₁′R₁′—CR₁′R₁′—,—NR₁′—C(O)—O—,

-   -   wherein        -   each R₁′ is independently selected from H or optionally            substituted C₁-C₃ straight or branched alkyl; and    -   R³² is selected from benzofuryl, methylfuryl, benzothienyl,        pyridyl, pyrazinyl, pyrimidinyl, pyrazolyl, wherein said        methylfuryl, pyridyl, pyrazinyl, pyrimidinyl or pyrazolyl is        optionally benzofused and wherein R³² is optionally substituted        or further substituted.

In a further aspect, the invention provides sirtuin-modulating compoundsof Structural Formula (XXII):

-   or a salt thereof, wherein:    -   R²¹ is selected from —NR₁′—C(O)—, —NR₁′—S(O)₂—,        —NR₁′—C(O)—NR₁′—, —NR₁′—C(S)—NR₁′—, —NR₁′—C(S)—NR₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—NR₁′—, —NR₁′—C(═NR₁′)—NR₁′—, —C(O)—NR₁′—,        —C(O)—NR₁′—S(O)₂—, —NR₁′—, —CR₁′R₁′—, —NR₁′—C(O)—CR₁′═CR₁′—,        —NR₁′—S(O)₂—NR₁′—, —NR₁′—C(O)—NR₁′—S(O)₂—,        —NR₁′—CR₁′R₁′—C(O)—NR₁′—, —CR₁′R₁′—C(O)—NR₁′—,        —NR₁′—C(O)—CR₁′═CR₁′—CR₁′R₁′—, —NR₁′—C(═N—CN)—NR₁′—,        —NR₁′—C(O)—CR₁′R₁′—, —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—O—,        —NR₁′—C(O)—CR₁′R₁′—O—, —NR₁′—S(O)₂—CR₁′R₁′—,        —NR₁′—S(O)₂—CR₁′R₁′—CR₁′R₁′—,

-   -    wherein each R₁′ is independently selected from H or optionally        substituted C₁-C₃ straight or branched alkyl; and    -   R³³ is an optionally substituted phenyl, wherein:    -   when R²¹ is —NR₁′—C(O)—, R₁′ is not H;    -   when R²¹ is —NH—C(O)—CH₂ or —NH—C(O)—CH₂—O—, R³³ is not        unsubstituted phenyl or 4-halophenyl; and    -   when R²¹ is —NH—S(O)₂—, R³³ is not unsubstituted phenyl, 2,4- or        3,4-dimethylphenyl, 2,4-dimethyl-5-methoxyphenyl,        2-methoxy-3,4-dichlorophenyl, 2-methoxy,        5-bromophenyl-3,4-dioxyethylenephenyl, 3,4-dimethoxyphenyl,        3,4-dichlorophenyl, 3,4-dimethylphenyl, 3- or 4-methylphenyl,        4-alkoxyphenyl, 4-phenoxyphenyl, 4-halophenyl, 4-biphenyl, or        4-acetylaminophenyl.

Preferably, R²¹ is selected from —NH—C(O)— or —NH—C(O)—CH₂—.

In one aspect, the invention provides sirtuin-modulating compounds ofStructural Formula (XXII):

-   or a salt thereof wherein:    -   R²¹ is selected from —NH—C(O)—, or —NH—C(O)—CH₂—; and    -   R³³ is phenyl substituted with    -   e) one —N(CH₃)₂ group;    -   f) one CN group at the 3 position;    -   g) one —S(CH₃) group; or

-   -    bridging the 3 and 4 positions.

In another aspect, the invention provides sirtuin-modulating compoundsof Structural Formula (XXIII):

-   or a salt thereof, wherein:    -   each R²⁰ and R^(20a) is independently selected from H or a        solubilizing group;    -   each R₁′, R₁″ and R₁′″ is independently selected from H or        optionally substituted C₁-C₃ straight or branched alkyl;    -   R²¹ is selected from —NR₁′—C(O)—, —NR₁′—S(O)₂—,        —NR₁′—C(O)—NR₁′—, —NR₁′—C(S)—NR₁′—, —NR₁′—C(S)—NR₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—NR₁′—, —NR₁′—C(═NR₁′)—NR₁′—,        —NR₁′—C(O)—CR₁′═CR₁′—, —NR₁′—S(O)₂—NR₁′—,        —NR₁′—C(O)—NR₁′—S(O)₂—, —NR₁′—CR₁′R₁′—C(O)—NR₁′—,        —NR₁′—C(O)—CR₁′═CR₁′—CR₁′R₁′—, —NR₁′—C(═N—CN)—NR₁′—,        —NR₁′—C(O)—CR₁′R₁′—O—, —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—O—,        —NR₁′—S(O)₂—CR₁′R₁′—, —NR₁′—S(O)₂—CR₁′R₁′—CR₁′R₁′—, or        —NR₁′—C(O)—CR₁′R₁′—; and    -   R³¹ is selected from an optionally substituted monocyclic or        bicyclic aryl, or an optionally substituted monocyclic or        bicyclic heteroaryl, with the provisos that:    -   when R²¹ is —NH—C(O)—, R³¹ is not is not 3,5-dinitrophenyl,        4-butoxyphenyl,

-   -   when R²¹ is —NH—C(O)— and each of R²⁰, R^(20a), R₁′, R₁″ and        R₁′″ is hydrogen, R³¹ is not

-   -    unsubstituted phenyl, 2- or 4-nitrophenyl, 2,4-dinitrophenyl,        2- or 4-chlorophenyl, 2-bromophenyl, 4-fluorophenyl,        2,4-dichlorophenyl, 2-carboxyphenyl, 2-azidophenyl, 2- or        4-aminophenyl, 2-acetamidophenyl, 4-methylphenyl, or        4-methoxyphenyl;    -   when R²¹ is —NH—C(O)—, R₁″ is methyl; and each of R²⁰, R^(20a),        R₁′ and R₁′″ is hydrogen, R³¹ is not 2-methylaminophenyl,

-   -   when R²¹ is —NH—C(O)—CH₂— or NH—C(S)—NH—, and each of R²⁰,        R^(20a), R₁′, R₁″ and R₁′″ is hydrogen, R³¹ is not unsubstituted        phenyl;    -   when R²¹ is —NH—S(O)₂—, R₁″ is hydrogen or methyl, and each of        R²⁰, R^(20a), R₁′ and R₁′″ is hydrogen, R³¹ is not        4-methylphenyl; and    -   when R²¹ is —NH—S(O)₂—, R^(20a) is hydrogen or —CH₂—N(CH₂CH₃)₂,        and each of R₂₀, R₁′, R₁″ and R₁′″ is hydrogen, R³¹ is not

In certain embodiments, R²¹ is selected from —NH—C(O)—, or—NH—C(O)—NR₁′—.

In certain embodiments, R³¹ is selected from optionally substitutedphenyl, quinoxalinyl or quinolinyl. For example, R³¹ is optionallysubstituted with up to 3 substituents independently selected from —OCH₃,—N(CH₃)₂, or a solubilizing group. Suitable examples of R³¹ include4-dimethylaminophenyl, 3,4-dimethoxyphenyl, 3,5-dimethoxyphenyl,3,4,5-trimethoxyphenyl, 3-methoxy-4-((piperazin-1-yl)methyl)phenyl,3-methoxy-4-((morpholino)methyl)phenyl,3-methoxy-4-((pyrrolidin-1-yl)methyl)phenyl, unsubstituted phenyl,unsubstituted quinoxalinyl, and unsubstituted quinolinyl.

In a particular aspect, the invention provides sirtuin-modulatingcompounds of Structural Formula (XXIII):

-   or a salt thereof, wherein:    -   each R²⁰ and R^(20a) is independently selected from H or a        solubilizing group;    -   each R₁′, R₁″ and R₁′″ is independently selected from H or        optionally substituted C₁-C₃ straight or branched alkyl;    -   R²¹ is selected from —NR₁′—C(O)—, —NR₁′—S(O)₂—,        —NR₁′—C(O)—NR₁′—, —NR₁′—C(S)—NR₁′—, —NR₁′—C(S)—NR₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—NR₁′—, —NR₁′—C(═NR₁′)—NR₁′—,        —NR₁′—C(O)—CR₁′═CR₁′—, —NR₁′—S(O)₂—NR₁′—,        —NR₁′—C(O)—NR₁′—S(O)₂—, —NR₁′—CR₁′R₁′—C(O)—NR₁′—,        —NR₁′—C(O)—CR₁′═CR₁′—CR₁′R₁′—, —NR₁′—C(═N—CN)—NR₁′—,        —NR₁′—C(O)—CR₁′R₁′—O—, —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—O—,        —NR₁′—S(O)₂—CR₁′R₁′—, —NR₁′—S(O)₂—CR₁′R₁′—CR₁′R₁′—, or        —NR₁′—C(O)—CR₁′R₁′—; and    -   R³¹ is selected from an optionally substituted monocyclic or        bicyclic aryl, or an optionally substituted monocyclic or        bicyclic heteroaryl,    -   wherein:    -   i) at least one R²⁰ is a solubilizing group or at least one R₁′″        is an optionally substituted C₁-C₃ straight or branched alkyl or        both; or    -   ii) R^(20a) is a solubilizing group other than CH₂—N(CH₂CH₃)₂.

In certain embodiments, R²¹ is selected from —NH—C(O)—, or—NH—C(O)—NR₁′—.

In certain embodiments, R³¹ is selected from optionally substitutedphenyl, quinoxalinyl or quinolinyl. For example, R³¹ is optionallysubstituted with up to 3 substituents independently selected from —OCH₃,—N(CH₃)₂, or a solubilizing group. Suitable examples of R³¹ include4-dimethylaminophenyl, 3,4-dimethoxyphenyl, 3,5-dimethoxyphenyl,3,4,5-trimethoxyphenyl, 3-methoxy-4-((piperazin-1-yl)methyl)phenyl,3-methoxy-4-((morpholino)methyl)phenyl,3-methoxy-4-((pyrrolidin-1-yl)methyl)phenyl, unsubstituted phenyl,unsubstituted quinoxalinyl, and unsubstituted quinolinyl.

In yet another aspect, the invention provides sirtuin-modulatingcompounds of Structural Formula (XXIV):

-   or a salt thereof, wherein:    -   each R²⁰ and R^(20a) is independently selected from H or a        solubilizing group;    -   each R₁′, R₁″ and R₁′″ is independently selected from H or        optionally substituted C₁-C₃ straight or branched alkyl;    -   R²¹ is selected from —NR²³—C(O)—, —NR₁′—S(O)₂—,        —NR₁′—C(O)—NR₁′—, —NR₁′—C(S)—NR₁′—, —NR₁′—C(S)—NR₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—NR₁′—, —NR₁′—C(═NR₁′)—NR₁′—,        —NR₁′—C(O)—CR₁′═CR₁′—, —NR₁′—S(O)₂—NR₁′—,        —NR₁′—C(O)—NR₁′—S(O)₂—, —NR₁′—CR₁′R₁′—C(O)—NR₁′—,        —NR₁′—C(O)—CR₁′═CR₁′—CR₁′R₁′—, —NR₁′—C(═N—CN)—NR₁′—,        —NR₁′—C(O)—CR₁′R₁′—O—, —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—O—,        —NR₁′—S(O)₂—CR₁′R₁′—, —NR₁′—S(O)₂—CR₁′R₁′—CR₁′R₁′—, or        —NR₁′—C(O)—CR₁′R₁′—; and    -   R³¹ is selected from an optionally substituted monocyclic or        bicyclic aryl, or an optionally substituted monocyclic or        bicyclic heteroaryl, with the provisos that:    -   when R²¹ is —NH—C(O)—CH₂—, R³¹ is not 2-methylphenyl, or        3,4-dimethoxyphenyl;    -   when R²¹ is —NH—C(O)—CH═CH—, R³¹ is not 2-chlorophenyl;    -   when R²¹ is —NH—C(O)—NH—, R³¹ is not unsubstituted        benzimidazolyl;    -   when R²¹ is —NH—S(O)₂—, and each of R²⁰, R^(20a), R₁′, R₁″ and        R₁′″ is hydrogen, R³¹ is not unsubstituted phenyl,        4-chlorophenyl, 4-methylphenyl, or 4-acetoamidophenyl;    -   when R²¹ is —NH—S(O)₂—, each of R₁′ and R₁′″ is methyl or        hydrogen, and each of R²⁰, R^(20a), and R₁″ is hydrogen, R³¹ is        not 4-nitrophenyl;    -   when R²¹ is —NH—C(O)—CH₂—O—, R₁′″ is methyl or hydrogen, and        each of R²⁰, R^(20a), R₁′, and R₁″ is hydrogen, R³¹ is not 2,3-,        2,5-, 2,6-, 3,4- or 3,5-dimethylphenyl, 2,4-dichloromethyl,        2,4-dimethyl-6-bromophenyl, 2- or 4-chlorophenyl,        2-(1-methylpropyl)phenyl, 5-methyl-2-(1-methylethyl)phenyl, 2-        or 4-methylphenyl, 2,4-dichloro-6-methylphenyl, nitrophenyl,        2,4-dimethyl-6-nitrophenyl, 2- or 4-methoxyphenyl,        4-acetyl-2-methoxyphenyl, 4-chloro-3,5-dimethylphenyl,        3-ethylphenyl, 4-bromophenyl, 4-cyclohexyphenyl,        4-(1-methylpropyl)phenyl, 4-(1-methylethyl)phenyl,        4-(1,1-dimethylethyl)phenyl, or unsubstituted phenyl;    -   when R²¹ is —NH—C(O)—CH₂—, R₁′″ is methyl or hydrogen, and each        of R²⁰, R^(20a), R₁′, and R₁″ is hydrogen, R³¹ is not        unsubstituted naphthyl, 4-chlorophenyl, 4-nitrophenyl,        4-methoxyphenyl, unsubstituted phenyl, unsubstituted thienyl

-   -   when R²¹ is —NH—C(O)—CH₂—, R₁′ is methyl, and each of R²⁰,        R^(20a), R₁″, and R₁′″ is hydrogen, R³¹ is not unsubstituted        phenyl;    -   when R²¹ is —NH—C(O)—CH═CH, R₁′″ is methyl or hydrogen, and each        of R²⁰, R^(20a), R₁′, and R₁″ is hydrogen, R³¹ is not        unsubstituted furyl, nitrophenyl-substituted furyl,        2,4-dichlorophenyl, 3,5-dichloro-2-methoxyphenyl, 3- or        4-nitrophenyl, 4-methoxyphenyl, unsubstituted phenyl, or        nitro-substituted thienyl;    -   when R²¹ is —NH—C(O)—CH(CH₂CH₃)—, and each of R²⁰, R^(20a), R₁′,        R₁″, and R₁′″ is hydrogen, R³¹ is not unsubstituted phenyl;    -   when R²¹ is —NH—C(O)—CH(CH₃)—O—, R₁′″ is methyl or hydrogen, and        each of R²⁰, R^(20a), R₁′, and R₁″ is hydrogen, R³¹ is not        2,4-dichlorophenyl.

In a particular aspect, the invention provides sirtuin-modulatingcompounds of Structural Formula (XXIV):

-   or a salt thereof, wherein:    -   each R²⁰ and R^(20a) is independently selected from H or a        solubilizing group and at least one of R²⁰ and R^(20a) is a        solubilizing group;    -   each R₁′, R₁″ and R₁′″ is independently selected from H or        optionally substituted C₁-C₃ straight or branched alkyl;    -   R²¹ is selected from —NR²³—C(O)—, —NR₁′—S(O)₂—,        —NR₁′—C(O)—NR₁′—, —NR₁′—C(S)—NR₁′—, —NR₁′—C(S)—NR₁′—CR₁′R₁′—,        —NR₁′—C(O)—CR₁′R₁′—NR₁′—, —NR₁′—C(═NR₁′)—NR₁′—,        —NR₁′—C(O)—CR₁′═CR₁′—, —NR₁′—S(O)₂—NR₁′—,        —NR₁′—C(O)—NR₁′—S(O)₂—, —NR₁′—CR₁′R₁′—C(O)—NR₁′—,        —NR₁′—C(O)—CR₁′═CR₁′—CR₁′R₁′, —NR₁′—C(═N—CN)—NR₁′—,        —NR₁′—C(O)—CR₁′R₁′—O—, —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—O—,        —NR₁′—S(O)₂—CR₁′R₁′—, —NR₁′—S(O)₂—CR₁′R₁′—CR₁′R₁′—, or        —NR₁′—C(O)—CR₁′R₁′—, wherein R²³ is an optionally substituted        C₁-C₃ straight or branched alkyl; and    -   R³¹ is selected from an optionally substituted monocyclic or        bicyclic aryl, or an optionally substituted monocyclic or        bicyclic heteroaryl.

In certain embodiments, when R²¹ is —NH—C(O)—CH₂—, R³¹ is not2-methylphenyl; or 3,4-dimethoxyphenyl; when R²¹ is —NH—C(O)—CH═CH—, R³¹is not 2-chlorophenyl; and/or when R²¹ is —NH—C(O)—NH—, R³¹ is notunsubstituted benzimidazolyl.

In a further aspect, the invention provides sirtuin-modulating compoundsof Structural Formula (XXV):

-   or a salt thereof, wherein:    -   each R²⁰ and R^(20a) is independently selected from H or a        solubilizing group, wherein at least one of R₂₀ and R^(20a) is a        solubilizing group;    -   each R₁′, R₁″ and R₁′″ is independently selected from H or        optionally substituted C₁-C₃ straight or branched alkyl; and    -   R³² is an optionally substituted phenyl.

In certain embodiments, R³² is selected from 3,4-dimethoxyphenyl,2,6-dimethoxyphenyl, or 2,4-dimethoxyphenyl; wherein R³² is furtheroptionally substituted with a solubilizing group.

In certain embodiments, R³² is not unsubstituted thienyl; unsubstitutedphenyl; 2-methylphenyl; 4-fluorophenyl; 4-methoxyphenyl; 4-methylphenyl;3,4-dioxyethylenephenyl; 3-acetylamino-4-methylphenyl;3-[(6-amino-1-oxohexyl)amino]-4-methylphenyl; 3-amino-4-methylphenyl;3,5-dimethoxyphenyl; 3-halo-4-methoxyphenyl; 3-nitro-4-methylphenyl; or4-propoxyphenyl.

In one aspect, the invention provides sirtuin-modulating compounds ofStructural Formula (XXVI):

-   or a salt thereof, wherein:    -   each R²⁰ and R^(20a) is independently selected from H or a        solubilizing group;    -   each R₁′, R₁″ and R₁′″ is independently selected from H or        optionally substituted C₁-C₃ straight or branched alkyl; and    -   R³³ is selected from an optionally substituted heteroaryl or an        optionally substituted bicyclic aryl, with the provisos that:    -   when each of R₁′ and R₁′″ is hydrogen or methyl and each of R₁″,        R₂₀ and R_(20a) is hydrogen, R³³ is not        5,6,7,8-tetrahydronaphthyl, unsubstituted benzofuryl,        unsubstituted benzothiazolyl, chloro- or nitro-substituted        benzothienyl, unsubstituted furyl, phenyl-, bromo- or        nitro-substituted furyl, dimethyl-substituted isoxazolyl,        unsubstituted naphthyl, 5-bromonaphthyl, 4-methylnaphthyl, 1- or        3-methoxynaphthyl, azo-substituted naphthyl, unsubstituted        pyrazinyl, S-methyl-substituted pyridyl, unsubstituted pyridyl,        thienyl- or phenyl-substituted quinolinyl, chloro-, bromo- or        nitro-substituted thienyl, unsubstituted thienyl, or

In a particular aspect, the invention provides sirtuin-modulatingcompounds of Structural Formula (XXVI):

-   -   or a salt thereof, wherein:        -   each R²⁰ and R^(20a) is independently selected from H or a            solubilizing group, wherein at least one of R²⁰ or R^(20a)            is a solubilizing group;        -   each R₁′, R₁″ and R₁′″ is independently selected from H or            optionally substituted C₁-C₃ straight or branched alkyl; and    -   R³³ is selected from an optionally substituted heteroaryl or an        optionally substituted bicyclic aryl.

In another aspect, the invention provides sirtuin-modulating compoundsof Structural Formula (XXVII):

-   -   wherein:        -   each R²⁰ and R^(20a) is independently selected from H or a            solubilizing group;        -   each R₁′ and R₁″ is independently selected from H or            optionally substituted C₁-C₃ straight or branched alkyl;    -   R¹⁹ is selected from:

-   -   wherein:        -   each Z₁₀, Z₁₁, Z₁₂ and Z₁₃ is independently selected from N,            CR²⁰, or CR₁′; and        -   each Z₁₄, Z₁₅ and Z₁₆ is independently selected from N,            NR₁′, S, O, CR²⁰, or CR₁′, wherein:        -   zero to two of Z₁₀, Z₁₁, Z₁₂ or Z₁₃ are N;        -   at least one of Z₁₄, Z₁₅ and Z₁₆ is N, NR₁′, S or O;        -   zero to one of Z₁₄, Z₁₅ and Z₁₆ is S or O;        -   zero to two of Z₁₄, Z₁₅ and Z₁₆ are N or NR₁′;        -   zero to one R²⁰ is a solubilizing group;        -   zero to one R₁′ is an optionally substituted C₁-C₃ straight            or branched alkyl; and        -   R²¹ is selected from —NR₁′—C(O)—, —NR₁′—S(O)₂—,            —NR₁′—C(O)—NR₁′—, —NR₁′—C(S)—NR₁′—,            —NR₁′—C(S)—NR₁′—CR₁′R₁′—, —NR₁′—C(O)—CR₁′R₁′—NR₁′—,            —NR₁′—C(═NR₁′)—NR₁′—, —C(O)—NR₁′—, —C(O)—NR₁′—S(O)₂—,            —NR₁′—, —CR₁′R₁′—, —NR₁′—C(O)—CR₁′═CR₁′—, —NR₁′—S(O)₂—NR₁′—,            —NR₁′—C(O)—NR₁′—S(O)₂—, —NR₁′—CR₁′R₁′—C(O)—NR₁′—,            —CR₁′R₁′—C(O)—NR₁′—, —NR₁′—C(O)—CR₁′═CR₁′—CR₁′R₁′—,            —NR₁′—C(═N—CN)—NR₁′—, —NR₁′—C(O)—CR₁′R₁′—O—,            —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—O—, —NR₁′—S(O)₂—CR₁′R₁′—,            —NR₁′—S(O)₂—CR₁′R₁′—CR₁′R₁′—, or —NR₁′—C(O)—CR₁′R₁′—; and    -   R³¹ is selected from an optionally substituted monocyclic or        bicyclic aryl, or an optionally substituted monocyclic or        bicyclic heteroaryl,        -   provided that when R²¹ is —NH—C(O)— and R¹⁹ is

-   -   -    R³¹ is not unsubstituted pyridyl, 2,6-dimethoxyphenyl,            3,4,5-trimethoxyphenyl or unsubstituted furyl.

In a particular aspect, the invention provides sirtuin-modulatingcompounds of Structural Formula (XXVII):

-   -   or a salt thereof, wherein:        -   each R²⁰ and R^(20a) is independently selected from H or a            solubilizing group;        -   each R₁′ and R₁″ is independently selected from H or            optionally substituted C₁-C₃ straight or branched alkyl;        -   R¹⁹ is selected from:

-   -   wherein:        -   each Z₁₀, Z₁₁, Z₁₂ and Z₁₃ is independently selected from N,            CR²⁰, or CR₁′; and        -   each Z₁₄, Z₁₅ and Z₁₆ is independently selected from N,            NR₁′, S, O, CR²⁰, or CR₁′, wherein:        -   zero to two of Z₁₀, Z₁₁, Z₁₂ or Z₁₃ are N;        -   at least one of Z₁₄, Z₁₅ and Z₁₆ is N, NR₁′, S or O;        -   zero to one of Z₁₄, Z₁₅ and Z₁₆ is S or O;        -   zero to two of Z₁₄, Z₁₅ and Z₁₆ are N or NR₁′;        -   zero to one R²⁰ is a solubilizing group;        -   zero to one R₁′ is an optionally substituted C₁-C₃ straight            or branched alkyl; and        -   R²¹ is selected from —NR₁′—C(O)—, —NR₁′—S(O)₂—,            —NR₁′—C(O)—NR₁′—, —NR₁′—C(S)—NR₁′—,            —NR₁′—C(S)—NR₁′—CR₁′R₁′—, —NR₁′—C(O)—CR₁′R₁′—NR₁′—,            —NR₁′—C(═NR₁′)—NR₁′—, —C(O)—NR₁′—, —C(O)—NR₁′—S(O)₂—,            —NR₁′—, —CR₁′R₁′—, —NR₁′—C(O)—CR₁′═CR₁′—, —NR₁′—S(O)₂—NR₁′—,            —NR₁′—C(O)—NR₁′—S(O)₂—, —NR₁′—CR₁′R₁′—C(O)—NR₁′—,            —CR₁′R₁′—C(O)—NR₁′—, —NR₁′—C(O)—CR₁′═CR₁′—CR₁′R₁′—,            —NR₁′—C(═N—CN)—NR₁′—, —NR₁′—C(O)—CR₁′R₁′—O—,            —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—O—, —NR₁′—S(O)₂—CR₁′R₁′—,            —NR₁′—S(O)₂—CR₁′R₁′—CR₁′R₁′—, or —NR₁′—C(O)—CR₁′R₁′—; and        -   R³¹ is selected from an optionally substituted monocyclic or            bicyclic aryl, or an optionally substituted monocyclic or            bicyclic heteroaryl, with the provisos that:        -   when R²¹ is —NH—C(O)—, R¹⁹ is not pyrazolyl;        -   when R²¹ is —NH—, and R¹⁹ is thiazolyl, R³¹ is not            optionally substituted phenyl or optionally substituted            pyridyl;        -   when R²¹ is —NH—C(O)—CH₂—, and R¹⁹ is pyrazolyl, R³¹ is not            unsubstituted indolyl or unsubstituted phenyl;        -   when R²¹ is —NH—C(O)—CH₂—, and R₁₉ is

-   -   -    R²¹ is not 2-methylphenyl or 3,4-dimethoxyphenyl;        -   when R²¹ is —NH—C(O)—CH═CH—, and R¹⁹ is

-   -   -    R³¹ is not 2-chlorophenyl;        -   when R²¹ is —NH—C(O)—NH—, and R¹⁹ is pyrazolyl, R³¹ is not            unsubstituted isoxazolyl, unsubstituted naphthyl,            unsubstituted phenyl, 2,6-difluorophenyl,            2,5-dimethylphenyl, 3,4-dichlorophenyl, or 4-chlorophenyl;        -   when R²¹ is —NH—C(O)—NH—, and R¹⁹ is

-   -   -    R³¹ is not unsubstituted benzimidazolyl;        -   when R²¹ is —NH—, and R¹⁹ is pyrazolyl, R³¹ is not            unsubstituted pyridyl;        -   when R^(20a) is a solubilizing group, R¹⁹ is            1-methylpyrrolyl and R²¹ is —NH—C(O)—, R³¹ is not            unsubstituted phenyl, unsubstituted furyl, unsubstituted            pyrrolyl, unsubstituted pyrazolyl, unsubstituted            isoquinolinyl, unsubstituted benzothienyl,            chloro-substituted benzothienyl, 2-fluoro-4-chlorophenyl or            phenyl singly substituted with a solubilizing group;

    -   when R^(20a) is a solubilizing group, R¹⁹ is thienyl and R²¹ is        —NH—C(O)—, R³¹ is not unsubstituted phenyl;

    -   when R^(20a) is a solubilizing group, R¹⁹ is methylimidazolyl        and R²¹ is —NH—C(O)—, R³¹ is not        1-methyl-4-(1,1-dimethylethyloxycarbonylamino)pyrrol-2-yl or        phenyl singly substituted with a solubilizing group;

    -   when R²¹ is —NH— and R¹⁹ is pyridyl, oxadiazolyl or        thiadiazolyl, R³¹ is not unsubstituted phenyl, 3-methoxyphenyl        or 4-methoxyphenyl;        -   when R²¹ is —NH—C(O)— and R¹⁹ is thiazolyl or pyrimidinyl,            R³¹ is not unsubstituted phenyl;        -   when R²¹ is —NH—C(O)— and R¹⁹ is

-   -   -    R³¹ is not unsubstituted pyridyl, unsubstituted thienyl,            unsubstituted phenyl, 2-methylphenyl, 4-fluorophenyl,            4-methoxyphenyl, 4-methylphenyl, 3,4-dioxyethylenephenyl,            3-acetylamino-4-methylphenyl,            3-[(6-amino-1-oxohexyl)amino]-4-methylphenyl,            3-amino-4-methylphenyl, 2,6-dimethoxyphenyl,            3,5-dimethoxyphenyl, 3-halo-4-methoxyphenyl,            3-nitro-4-methylphenyl, 4-propoxyphenyl,            3,4,5-trimethoxyphenyl or unsubstituted furyl;

    -   when R²¹ is —NH—C(O)— and R¹⁹ is

-   -    R³¹ is not 3,5-dinitrophenyl, 4-butoxyphenyl,

In certain embodiments, R²¹ is selected from —NH—C(O)— or—NH—C(O)—NR₁′—, preferably —NH—C(O)—.

In certain embodiments, R³¹ is selected from optionally substitutedphenyl, quinoxalinyl or quinolinyl; preferably optionally substitutedphenyl. For example, R³¹ is optionally substituted with up to 3substituents independently selected from —OCH₃, —N(CH₃)₂, or asolubilizing group. Suitable examples of R³¹ include4-dimethylaminophenyl; 3,4-dimethoxyphenyl; 3,5-dimethoxyphenyl;3,4,5-trimethoxyphenyl; 3-methoxy-4-((piperazin-1-yl)methyl)phenyl;3-methoxy-4-((morpholino)methyl)phenyl;3-methoxy-4-((pyrrolidin-1-yl)methyl)phenyl; unsubstituted phenyl;unsubstituted quinoxalinyl; and unsubstituted quinolinyl. Preferredexamples of R³¹ include 3,4-dimethoxyphenyl; 2,6-dimethoxyphenyl; or2,4-dimethoxyphenyl; wherein R³¹ is further optionally substituted witha solubilizing group.

In preferred embodiments, R²¹ is —NH—C(O)— and R³¹ is selected from3-methoxyphenyl; 3,4-dimethoxyphenyl; 3,4,5-trimethoxyphenyl; or4-dimethylaminophenyl.

In certain embodiments, when R²¹ is —NH—C(O)—, R¹⁹ is not

In certain embodiments, when R²¹ is —NH—C(O)—, R¹⁹ is not optionallysubstituted pyrazolyl, thiazolyl, thienyl, pyrrolyl or pyrimidinyl; whenR²¹ is —NH—C(O)—CH2- or —NH—C(O)—NH—, R¹⁹ is not pyrazolyl; and/or whenR²¹ is —NH—, R¹⁹ is not optionally substituted pyridyl, thiazolyl,pyrazolyl, thiadiazolyl, or oxadiazolyl.

In a more particular aspect, the invention provides sirtuin-modulatingcompounds of Structural Formula (XXVII):

-   -   or a salt thereof, wherein:        -   each R²⁰ and R^(20a) is independently selected from H or a            solubilizing group;    -   each R₁′ and R₁″ is independently selected from H or optionally        substituted C₁-C₃ straight or branched alkyl;    -   R¹⁹ is selected from:

-   -   wherein:        -   each Z₁₀, Z₁₁, Z₁₂ and Z₁₃ is independently selected from N,            CR²⁰, or CR₁′; and        -   each Z₁₄, Z₁₅ and Z₁₆ is independently selected from N,            NR₁′, S, O, CR²⁰, or CR₁′, wherein:        -   one to two of Z₁₀, Z₁₁, Z₁₂ or Z₁₃ are N;        -   at least one of Z₁₄, Z₁₅ and Z₁₆ is N, NR₁′, S or O;        -   zero to one of Z₁₄, Z₁₅ and Z₁₆ is S or O;        -   zero to two of Z₁₄, Z₁₅ and Z₁₆ are N or NR₁′;        -   zero to one R²⁰ is a solubilizing group;        -   zero to one R₁′″ is an optionally substituted C₁-C₃ straight            or branched alkyl; and        -   R²¹ is selected from —NR₁′—C(O)—, —NR₁′—S(O)₂—,            —NR₁′—C(O)—NR₁′—, —NR₁′—C(S)—NR₁′—,            —NR₁′—C(S)—NR₁′—CR₁′R₁′—, —NR₁′—C(O)—CR₁′R₁′—NR₁′—,            —NR₁′—C(═NR₁′)—NR₁′—, —NR₁′—C(O)—CR₁′═CR₁′—,            —NR₁′—S(O)₂—NR₁′—, —NR₁′—C(O)—NR₁′—S(O)₂—,            —NR₁′—CR₁′R₁′—C(O)—NR₁′—, —NR₁′—C(O)—CR₁′═CR₁′—CR₁′R₁′—,            —NR₁′—C(═N—CN)—NR₁′—, —NR₁′—C(O)—CR₁′R₁′—O—,            —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—O—, —NR₁′—S(O)₂—CR₁′R₁′—,            —NR₁′—S(O)₂—CR₁′R₁′—CR₁′R₁′—, or —NR₁′—C(O)—CR₁′R₁′—; and        -   R³¹ is selected from an optionally substituted monocyclic or            bicyclic aryl, or an optionally substituted monocyclic or            bicyclic heteroaryl, with the provisos that:        -   when R²¹ is —NH—C(O)—, R¹⁹ is not pyrazolyl;        -   when R²¹ is —NH—C(O)—CH₂—, and R¹⁹ is pyrazolyl, R³¹ is not            unsubstituted indolyl or unsubstituted phenyl;        -   when R²¹ is —NH—C(O)—NH—, and R¹⁹ is pyrazolyl, R³¹ is not            unsubstituted isoxazolyl, unsubstituted naphthyl,            unsubstituted phenyl, 2,6-difluorophenyl;            2,5-dimethylphenyl; 3,4-dichlorophenyl; or 4-chlorophenyl;    -   when R^(20a) is a solubilizing group, R¹⁹ is 1-methylpyrrolyl        and R²¹ is —NH—C(O)—, R³¹ is not unsubstituted phenyl;        unsubstituted furyl; unsubstituted pyrrolyl; unsubstituted        pyrazolyl; unsubstituted isoquinolinyl; unsubstituted        benzothienyl; chloro-substituted benzothienyl;        2-fluoro-4-chlorophenyl or phenyl singly substituted with a        solubilizing group;    -   when R^(20a) is a solubilizing group, R¹⁹ is thienyl and R²¹ is        —NH—C(O)—, R³¹ is not unsubstituted phenyl;        -   when R^(20a) is a solubilizing group, R¹⁹ is            methylimidazolyl and R²¹ is —NH—C(O)—, R³¹ is not            1-methyl-4-(1,1-dimethylethyloxycarbonylamino)pyrrol-2-yl or            phenyl singly substituted with a solubilizing group; and        -   when R²¹ is —NH—C(O)— and R¹⁹ is thiazolyl or pyrimidinyl,            R³¹ is not unsubstituted phenyl.

In certain embodiments, R²¹ is selected from —NH—C(O)— or—NH—C(O)—NR₁′—, preferably —NH—C(O)—.

In certain embodiments, R³¹ is selected from optionally substitutedphenyl, quinoxalinyl or quinolinyl; preferably optionally substitutedphenyl. For example, R³¹ is optionally substituted with up to 3substituents independently selected from —OCH₃, —N(CH₃)₂, or asolubilizing group. Suitable examples of R³¹ include4-dimethylaminophenyl; 3,4-dimethoxyphenyl; 3,5-dimethoxyphenyl;3,4,5-trimethoxyphenyl; 3-methoxy-4-((piperazin-1-yl)methyl)phenyl;3-methoxy-4-((morpholino)methyl)phenyl;3-methoxy-4-((pyrrolidin-1-yl)methyl)phenyl; unsubstituted phenyl;unsubstituted quinoxalinyl; and unsubstituted quinolinyl. Preferredexamples of R³¹ include 3,4-dimethoxyphenyl; 2,6-dimethoxyphenyl; or2,4-dimethoxyphenyl; wherein R³¹ is further optionally substituted witha solubilizing group.

-   -   In preferred embodiments, R²¹ is —NH—C(O)— and R³¹ is selected        from 3-methoxyphenyl; 3,4-dimethoxyphenyl;        3,4,5-trimethoxyphenyl; or 4-dimethylaminophenyl.

In yet another aspect, the invention provides compounds of StructuralFormula (XXVIII):

-   -   or a salt thereof, wherein:        -   each R²⁰ and R^(20a) is independently selected from H or a            solubilizing group;        -   each R₁′ and R₁″ is independently selected from H or            optionally substituted C₁-C₃ straight or branched alkyl;        -   R²⁹ is selected from:

-   -   wherein:        -   each Z₁₀, Z₁₁, Z₁₂ and Z₁₃ is independently selected from N,            CR²⁰, or CR₁′, wherein one of Z₁₀, Z₁₁, Z₁₂ or Z₁₃ is N; and        -   zero to one R²⁰ is a solubilizing group;        -   zero to one R₁′″ is an optionally substituted C₁-C₃ straight            or branched alkyl; and        -   R²¹ is selected from —NR₁′—C(O)—, —NR₁′—S(O)₂—,            —NR₁′—C(O)—NR₁′—, —NR₁′—C(S)—NR₁′—,            —NR₁′—C(S)—NR₁′—CR₁′R₁′—, —NR₁′—C(O)—CR₁′R₁′—NR₁′—,            —NR₁′—C(═NR₁′)—NR₁′—, —NR₁′—C(O)—CR₁′═CR₁′—,            —NR₁′—S(O)₂—NR₁′—, —NR₁′—C(O)—NR₁′—S(O)₂—,            —NR₁′—CR₁′R₁′—C(O)—NR₁′, —NR₁′—C(O)—CR₁′═CR₁′—CR₁′R₁′—,            —NR₁′—C(═N—CN)—NR₁′—, —NR₁′—C(O)—CR₁′R₁′—O—,            —NR₁′—C(O)—CR₁′R₁′—CR₁′R₁′—O—, —NR₁′—S(O)₂—CR₁′R₁′—,            —NR₁′—S(O)₂—CR₁′R₁′—CR₁′R₁′—, or —NR₁′—C(O)—CR₁′R₁′—; and        -   R³¹ is selected from an optionally substituted monocyclic or            bicyclic aryl, or an optionally substituted monocyclic or            bicyclic heteroaryl.

In certain embodiments, R³¹ is optionally substituted phenyl, such as3-methoxyphenyl, 3,4-dimethoxyphenyl, 3,4,5-trimethoxyphenyl, or4-dimethylaminophenyl.

In certain embodiments, R²¹ is —NH—C(O)—.

In preferred embodiments, R²¹ is —NH—C(O)— and R³¹ is an optionallysubstituted phenyl, such as 3-methoxyphenyl, 3,4-dimethoxyphenyl,3,4,5-trimethoxyphenyl, or 4-dimethylaminophenyl.

In a further aspect, such as when the sirtuin modulator is a sirtuininhibitor, the invention provides novel sirtuin-modulating compounds ofFormula (VI):

-   or a salt thereof, wherein:    -   Het is an optionally substituted heterocyclic aryl group; and    -   Ar′ is an optionally substituted carbocyclic or heterocyclic        aryl group.

In certain embodiments, Het comprises one N heteroatom and 1 to 2additional heteroatoms independently selected from N, O or S, such asoxazolopyridyl.

In certain embodiments, Ar′ is selected from optionally substitutedphenyl, benzothiazolyl, or benzoxazolyl. When Ar′ is substituted phenyl,typically it is substituted with 1 to 3 substituents independentlyselected from halo, methyl, O-methyl, S-methyl or N(CH₃)₂, morpholino,or 3,4 dioxymethylene.

Compounds of the invention, including novel compounds of the invention,can also be used in the methods described herein.

The compounds and salts thereof described herein also include theircorresponding hydrates (e.g., hemihydrate, monohydrate, dihydrate,trihydrate, tetrahydrate) and solvates. Suitable solvents forpreparation of solvates and hydrates can generally be selected by askilled artisan.

The compounds and salts thereof can be present in amorphous orcrystalline (including co-crystalline and polymorph) forms.

In the compounds described above, bivalent groups disclosed as possiblevalues for variables can have either orientation, provided that suchorientation results in a stable molecule. Preferably, however, the lefthand side of a bivalent group (e.g., —NR₁′—C(O)—) is attached to abivalent arylene or heteroarylene group (e.g., R¹⁹) and the right handside of a bivalent group is attached to a monovalent aryl group (e.g.,R³¹).

Sirtuin-modulating compounds of the invention having hydroxylsubstituents, unless otherwise indicated, also include the relatedsecondary metabolites, such as phosphate, sulfate, acyl (e.g., acetyl,fatty acid acyl) and sugar (e.g., glucurondate, glucose) derivatives(e.g., of hydroxyl groups), particularly the sulfate, acyl and sugarderivatives. In other words, substituent groups —OH also include —OSO₃^(− M) ⁺, where M⁺ is a suitable cation (preferably H⁺, NH₄ ⁺ or analkali metal ion such as Na⁺ or K⁺) and sugars such as

These groups are generally cleavable to —OH by hydrolysis or bymetabolic (e.g., enzymatic) cleavage.

In certain embodiments, the compounds of the invention exclude one ormore of the species disclosed in Tables 4-6. In certain suchembodiments, the compounds of the invention exclude compound 7.

Sirtuin-modulating compounds of the invention advantageously modulatethe level and/or activity of a sirtuin protein, particularly thedeacetylase activity of the sirtuin protein.

Separately or in addition to the above properties, certainsirtuin-modulating compounds of the invention do not substantially haveone or more of the following activities: inhibition of PI3-kinase,inhibition of aldoreductase, inhibition of tyrosine kinase,transactivation of EGFR tyrosine kinase, coronary dilation, orspasmolytic activity, at concentrations of the compound that areeffective for modulating the deacetylation activity of a sirtuin protein(e.g., such as a SIRT1 and/or a SIRT3 protein).

An alkyl group is a straight chained, branched or cyclic non-aromatichydrocarbon which is completely saturated. Typically, a straight chainedor branched alkyl group has from 1 to about 20 carbon atoms, preferablyfrom 1 to about 10, and a cyclic alkyl group has from 3 to about 10carbon atoms, preferably from 3 to about 8. Examples of straight chainedand branched alkyl groups include methyl, ethyl, n-propyl, iso-propyl,n-butyl, sec-butyl, tert-butyl, pentyl, hexyl, pentyl and octyl. A C1-C4straight chained or branched alkyl group is also referred to as a “loweralkyl” group.

An alkenyl group is a straight chained, branched or cyclic non-aromatichydrocarbon which contains one or more double bonds. Typically, thedouble bonds are not located at the terminus of the alkenyl group, suchthat the double bond is not adjacent to another functional group.

An alkynyl group is a straight chained, branched or cyclic non-aromatichydrocarbon which contains one or more triple bonds. Typically, thetriple bonds are not located at the terminus of the alkynyl group, suchthat the triple bond is not adjacent to another functional group.

A ring (e.g., 5- to 7-membered ring) or cyclic group includescarbocyclic and heterocyclic rings. Such rings can be saturated orunsaturated, including aromatic. Heterocyclic rings typically contain 1to 4 heteroatoms, although oxygen and sulfur atoms cannot be adjacent toeach other.

Aromatic (aryl) groups include carbocyclic aromatic groups such asphenyl, naphthyl, and anthracyl, and heteroaryl groups such asimidazolyl, thienyl, furyl, pyridyl, pyrimidyl, pyranyl, pyrazolyl,pyrroyl, pyrazinyl, thiazolyl, oxazolyl, and tetrazolyl.

Aromatic groups also include fused polycyclic aromatic ring systems inwhich a carbocyclic aromatic ring or heteroaryl ring is fused to one ormore other heteroaryl rings. Examples include benzothienyl, benzofuryl,indolyl, quinolinyl, benzothiazole, benzoxazole, benzimidazole,quinolinyl, isoquinolinyl and isoindolyl.

Non-aromatic heterocyclic rings are non-aromatic carbocyclic rings whichinclude one or more heteroatoms such as nitrogen, oxygen or sulfur inthe ring. The ring can be five, six, seven or eight-membered. Examplesinclude tetrahydrofuryl, tetrahyrothiophenyl, morpholino,thiomorpholino, pyrrolidinyl, piperazinyl, piperidinyl, andthiazolidinyl, along with the cyclic form of sugars.

A ring fused to a second ring shares at least one common bond.

Suitable substituents on an alkyl, alkenyl, alkynyl, aryl, non-aromaticheterocyclic or aryl group (carbocyclic and heteroaryl) are those whichdo not substantially interfere with the ability of the disclosedcompounds to have one or more of the properties disclosed herein. Asubstituent substantially interferes with the properties of a compoundwhen the magnitude of the property is reduced by more than about 50% ina compound with the substituent compared with a compound without thesubstituent. Examples of suitable substituents include —OH, halogen(—Br, —Cl, —I and —F), —OR^(a), —O—COR^(a), —COR^(a), —C(O)R^(a), —CN,—NO², —COOH, —COOR^(a), —OCO₂R^(a), —C(O)NR^(a)R^(b), —OC(O)NR^(a)R^(b),—SO₃H, —NH₂, —NHR^(a), —N(R^(a)R^(b)), —COOR^(a), —CHO, —CONH₂,—CONHR^(a), —CON(R^(a)R^(b)), —NHCOR^(a), —NRCOR^(a), —NHCONH₂,—NHCONR^(a)H, —NHCON(R^(a)R^(b)), —NR^(c)CONH₂, —NR^(c)CONR^(a)H,—NR^(c)CON(R^(a)R^(b)), —C(═NH)—NH₂, —C(═NH)—NHR^(a),—C(═NH)—N(R^(a)R^(b)), —C(═NR^(c))—NH₂, —C(═NR^(c))—NHR^(a),—C(═NR^(c))—N(R^(a)R^(b)), —NH—C(═NH)—NH₂, —NH—C(═NH)—NHR^(a),—NH—C(═NH)—N(R^(a)R^(b)), —NH—C(═NR^(c))—NH₂, —NH—C(═NR^(c))—NHR^(a),—NH—C(═NR^(c))—N(R^(a)R^(b)), NR^(d)—C(═NH)—NH₂, —NR^(d)—C(═NH)—NHR^(a),—NR^(d)—C(═NH)—N(R^(a)R^(b)), —NR^(d)C(═NR^(c))—NH₂,—NR^(d)—C(═NR^(c))—NHR^(a), —NR^(d)—C(═NR^(c))—N(R^(a)R^(b)), —NHNH₂,—NHNHR^(a), —NHR^(a)R^(b), —SO₂NH₂, —SO₂NHR^(a), —SO₂NR^(a)R^(b),—CH═CHR^(a), —CH═CR^(a)R^(b), —CR^(c)═CR^(a)R^(b), CR^(c)═CHR^(a),—CR^(c)═CR^(a)R^(b), —CCR^(a), —SH, —SO_(k)R^(a) (k is 0, 1 or 2),—S(O)_(k)OR^(a) (k is 0, 1 or 2) and —NH—C(═NH)—NH₂. R^(a)—R^(d) areeach independently an aliphatic, substituted aliphatic, benzyl,substituted benzyl, aromatic or substituted aromatic group, preferablyan alkyl, benzylic or aryl group. In addition, —NR^(a)R^(b), takentogether, can also form a substituted or unsubstituted non-aromaticheterocyclic group. A non-aromatic heterocyclic group, benzylic group oraryl group can also have an aliphatic or substituted aliphatic group asa substituent. A substituted aliphatic group can also have anon-aromatic heterocyclic ring, a substituted a non-aromaticheterocyclic ring, benzyl, substituted benzyl, aryl or substituted arylgroup as a substituent. A substituted aliphatic, non-aromaticheterocyclic group, substituted aryl, or substituted benzyl group canhave more than one substituent.

Combinations of substituents and variables envisioned by this inventionare only those that result in the formation of stable compounds. As usedherein, the term “stable” refers to compounds that possess stabilitysufficient to allow manufacture and that maintain the integrity of thecompound for a sufficient period of time to be useful for the purposesdetailed herein.

A hydrogen-bond donating group is a functional group having a partiallypositively-charged hydrogen atom (e.g., —OH, —NH₂, —SH) or a group(e.g., an ester) that metabolizes into a group capable of donating ahydrogen bond.

As used herein, a “solubilizing group” is a moiety that has hydrophiliccharacter sufficient to improve or increase the water-solubility of thecompound in which it is included, as compared to an analog compound thatdoes not include the group. The hydrophilic character can be achieved byany means, such as by the inclusion of functional groups that ionizeunder the conditions of use to form charged moieties (e.g., carboxylicacids, sulfonic acids, phosphoric acids, amines, etc.); groups thatinclude permanent charges (e.g., quaternary ammonium groups); and/orheteroatoms or heteroatomic groups (e.g., O, S, N, NH,N—(CH₂)_(y)—R^(a), N—(CH₂)_(y)—C(O)R^(a), N—(CH₂)_(y)—C(O)OR^(a),N—(CH₂)_(y)—S(O)₂R^(a)—, N—(CH₂)_(y)—S(O)₂OR^(a),N—(CH₂)_(y)—C(O)NR^(a)R^(a), etc., wherein R^(a) is selected fromhydrogen, lower alkyl, lower cycloalkyl, (C6-C14) aryl, phenyl,naphthyl, (C7-C20) arylalkyl and benzyl, wherein R^(a) is optionallysubstituted; and y is an integer ranging from 0 to 6), optionallysubstituted heterocyclic groups (e.g., —(CH₂)_(n)—R^(b),—(CH₂)_(n)—C(O)—R^(b), —(CH₂)_(n)—O—(CH₂)_(n)—R^(b), wherein R^(b) isselected from an optionally substituted saturated monocyclicheterocycle, an optionally substituted saturated bicyclic fusedheterocycle, an optionally substituted saturated bicyclic spiroheterocycle, an optionally substituted heteroaryl and an optionallysubstituted partially substituted non-aryl heterocycle; and n is aninteger ranging from 0 to 2). It should be understood that substituentspresent on R^(a) or R^(b) need not improve or increase water solubilityover their unsubstituted counterparts to be within the scope of thisdefinition. All that is required is that such substituents do notsignificantly reverse the improvement in water-solubility afforded bythe unsubstituted R^(a) or R^(b) moiety.

In one embodiment, the solubilizing group increases the water-solubilityof the corresponding compound lacking the solubilizing group at least5-fold, preferably at least 10-fold, more preferably at least 20-foldand most preferably at least 50-fold.

In one preferred embodiment, the solubilizing group is a moiety of theformula: —(CH₂)_(n)—R¹⁰⁰—N(R¹⁰¹)(R¹⁰¹), wherein:

-   n is selected from 0, 1 or 2;-   R¹⁰⁰ is selected from a bond, —C(O)—, or —O(CH₂)_(n); and-   each R¹⁰¹ is independently selected from:    -   a. hydrogen;    -   b. C₁-C₄ straight or branched alkyl, wherein said alkyl is        optionally substituted with halo, CN, OH, O—(C₁-C₄ straight or        branched alkyl), N(R₁′)(R₁′), or ═O;

-   -   f. both R¹⁰¹ moieties are taken together with the nitrogen atom        to which they are bound to form a ring of the structure

-   -   g. both R¹⁰¹ moieties are taken together with the nitrogen atom        to which they are bound to form a 5-membered heteroaryl ring        containing 1 to 3 additional N atoms, wherein said heteroaryl        ring is optionally substituted with R₁′;

-   wherein:    -   each Z is independently selected from —O—, —S—, —NR₁′—, or        —C(R⁵⁰)(R⁵⁰)—, wherein:        -   at least three of Z₂₀, Z₂₁, Z₂₂, and Z₂₃ are —C(R⁵⁰)(R⁵⁰)—;        -   at least three of Z₂₄, Z₂₅, Z₂₆, Z₂₇, and Z₂₈ are            —C(R⁵⁰)(R⁵⁰)—;        -   at least four of Z₃₀, Z₃₁, Z₃₂, and Z₃₃ are —C(R⁵⁰)(R⁵⁰)—;            and        -   at least four of Z₃₄, Z₃₅, Z₃₆, Z₃₇, and Z₃₈ are            —C(R⁵⁰)(R⁵⁰)—;    -   each R₁′ is independently selected from hydrogen or a C₁-C₃        straight or branched alkyl optionally substituted with one or        more substituent independently selected from halo, —CN, —OH,        —OCH₃, —NH₂, —NH(CH₃), —N(CH₃)₂, or ═O;    -   each R⁵⁰ is independently selected from R₁′, halo, CN, OH,        O—(C₁-C₄ straight or branched alkyl), N(R₁′)(R₁′), ═CR₁′, SR₁′,        ═NR₁′, ═NOR₁′, or ═O;    -   any two suitable non-cyclic R⁵⁰ are optionally bound to one        another directly or via a C₁ to C₂ alkylene, alkenylene or        alkanediylidene bridge to produce a bicyclic fused or spiro        ring; and

ring structure is optionally benzofused or fused to a monocyclicheteroaryl to produce a bicyclic ring.

For clarity, the term “C₁ to C₂ alkylene, alkenylene or alkanediylidenebridge” means the multivalent structures —CH₂—, —CH₂—CH₂—, —CH═, ═CH—,—CH═CH—, or ═CH—CH═. The two R⁵⁰ moieties that are optionally bound toone another can be either on the same carbon atom or different carbonatoms. The former produces a spiro bicyclic ring, while the latterproduces a fused bicyclic ring. It will be obvious to those of skill inthe art that when two R⁵⁰ are bound to one another to form a ring(whether directly or through one of the recited bridges), one or moreterminal hydrogen atoms on each R⁵⁰ will be lost. Accordingly, a“suitable non-cyclic R⁵⁰” moiety available for forming a ring is anon-cyclic R⁵⁰ that comprises at least one terminal hydrogen atom.

In another preferred embodiment, the solubilizing group is a moiety ofthe formula: —(CH₂)_(n)—O—R¹⁰¹, wherein n and R¹⁰¹ are as defined above.

In another preferred embodiment, the solubilizing group is a moiety ofthe formula: —(CH₂)_(n)—C(O)—R₁′, wherein n and R₁′ are as definedabove.

In a more preferred embodiment, a solubilizing group is selected from—(CH₂)_(n)—R₁₀₂, wherein n is 0, 1 or 2; and R₁₀₂ is selected from

wherein R₁′ are as defined above.

In an even more preferred embodiment, a solubilizing group is selectedfrom 2-dimethylaminoethylcarbamoyl, piperazin-1-ylcarbonyl,piperazinylmethyl, dimethylaminomethyl, 4-methylpiperazin-1-ylmethyl,4-aminopiperidin-1-yl-methyl, 4-fluoropiperidin-1-yl-methyl,morpholinomethyl, pyrrolidin-1-ylmethyl,2-oxo-4-benzylpiperazin-1-ylmethyl, 4-benzylpiperazin-1-ylmethyl,3-oxopiperazin-1-ylmethyl, piperidin-1-ylmethyl, piperazin-1-ylethyl,2,3-dioxopropylaminomethyl, thiazolidin-3-ylmethyl,4-acetylpiperazin-1-ylmethyl, 4-acetylpiperazin-1-yl, morpholino,3,3-difluoroazetidin-1-ylmethyl, 2H-tetrazol-5-ylmethyl,thiomorpholin-4-ylmethyl, 1-oxothiomorpholin-4-ylmethyl,1,1-dioxothiomorpholin-4-ylmethyl, 1H-imidazol-1-ylmethyl,3,5-dimethylpiperazin-1ylmethyl, 4-hydroxypiperidin-1-ylmethyl,N-methyl(1-acetylpiperidin-4-yl)-aminomethyl,N-methylquinuclidin-3-ylaminomethyl, 1H-1,2,4-triazol-1-ylmethyl,1-methylpiperidin-3-yl-oxymethyl, or 4-fluoropiperidin-1-yl.

To the extent not included within any of the definitions set forthabove, the term “solubilizing group” also includes moieties disclosed asbeing attached to the 7-position of1-cyclopropyl-6-fluoro-1,4-dihydro-4-oxoquinoline-3-carboxylic acid(ciprofloxacin) and its derivatives, as disclosed in PCT publications WO2005026165, WO 2005049602, and WO 2005033108, and European Patentpublications EP 0343524, EP 0688772, EP 0153163, EP 0159174; as well as“water-solubilizing groups” described in United States patentpublication 2006/0035891. The disclosure of each of these patentpublications is incorporated herein by reference.

Double bonds indicated in a structure as:

are intended to include both the (E)- and (Z)-configuration. Preferably,double bonds are in the (E)-configuration.

A sugar is an aldehyde or ketone derivative of a straight-chainpolyhydroxy alcohol, which contains at least three carbon atoms. A sugarcan exist as a linear molecule or, preferably, as a cyclic molecule(e.g., in the pyranose or furanose form). Preferably, a sugar is amonosaccharide such as glucose or glucuronic acid. In embodiments of theinvention where, for example, prolonged residence of a compoundderivatized with a sugar is desired, the sugar is preferably anon-naturally occurring sugar. For example, one or more hydroxyl groupsare substituted with another group, such as a halogen (e.g., chlorine).The stereochemical configuration at one or more carbon atoms can also bealtered, as compared to a naturally occurring sugar. One example of asuitable non-naturally occurring sugar is sucralose.

A fatty acid is a carboxylic acid having a long-chained hydrocarbonmoiety. Typically, a fatty acid has an even number of carbon atomsranging from 12 to 24, often from 14 to 20. Fatty acids can be saturatedor unsaturated and substituted or unsubstituted, but are typicallyunsubstituted. Fatty acids can be naturally or non-naturally occurring.In embodiments of the invention where, for example, prolonged residencetime of a compound having a fatty acid moiety is desired, the fatty acidis preferably non-naturally occurring. The acyl group of a fatty acidconsists of the hydrocarbon moiety and the carbonyl moiety of thecarboxylic acid functionality, but excludes the —OH moiety associatedwith the carboxylic acid functionality.

Also included in the present invention are salts, particularlypharmaceutically acceptable salts, of the sirtuin-modulating compoundsdescribed herein. The compounds of the present invention that possess asufficiently acidic, a sufficiently basic, or both functional groups,can react with any of a number of inorganic bases, and inorganic andorganic acids, to form a salt. Alternatively, compounds that areinherently charged, such as those with a quaternary nitrogen, can form asalt with an appropriate counterion (e.g., a halide such as bromide,chloride, or fluoride, particularly bromide).

Acids commonly employed to form acid addition salts are inorganic acidssuch as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuricacid, phosphoric acid, and the like, and organic acids such asp-toluenesulfonic acid, methanesulfonic acid, oxalic acid,p-bromophenyl-sulfonic acid, carbonic acid, succinic acid, citric acid,benzoic acid, acetic acid, and the like. Examples of such salts includethe sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate,monohydrogenphosphate, dihydrogenphosphate, metaphosphate,pyrophosphate, chloride, bromide, iodide, acetate, propionate,decanoate, caprylate, acrylate, formate, isobutyrate, caproate,heptanoate, propiolate, oxalate, malonate, succinate, suberate,sebacate, fumarate, maleate, butyne-1,4-dioate, hexyne-1,6-dioate,benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate,hydroxybenzoate, methoxybenzoate, phthalate, sulfonate, xylenesulfonate,phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate,gamma-hydroxybutyrate, glycolate, tartrate, methanesulfonate,propanesulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate,mandelate, and the like.

Base addition salts include those derived from inorganic bases, such asammonium or alkali or alkaline earth metal hydroxides, carbonates,bicarbonates, and the like. Such bases useful in preparing the salts ofthis invention thus include sodium hydroxide, potassium hydroxide,ammonium hydroxide, potassium carbonate, and the like.

According to another embodiment, the present invention provides methodsof producing the above-defined sirtuin-modulating compounds. Thecompounds may be synthesized using conventional techniques.Advantageously, these compounds are conveniently synthesized fromreadily available starting materials.

Thus, one embodiment relates to a method of making a compound of thestructure described herein using the following synthesis scheme:

One of skill in the art would recognize that this synthetic scheme, orsimilar variants, usefully allows the incorporation of a variety of Rgroups into compounds falling within the scope of the instant invention,for example, compounds of the tables below.

As can be appreciated by the skilled artisan, the above synthetic schemeis not intended to comprise a comprehensive list of all means by whichthe compounds described and claimed in this application may besynthesized. Further methods will be evident to those of ordinary skillin the art. Additionally, the various synthetic steps described abovemay be performed in an alternate sequence or order to give the desiredcompounds. Synthetic chemistry transformations and methodologies usefulin synthesizing the sirtuin-modulating compounds described herein areknown in the art and include, for example, those described in R. Larock,Comprehensive Organic Transformations (1989); T. W. Greene and P. G. M.Wuts, Protective Groups in Organic Synthesis, 2d. Ed. (1991); L. Fieserand M. Fieser, Fieser and Fieser's Reagents for Organic Synthesis(1994); and L. Paquette, ed., Encyclopedia of Reagents for OrganicSynthesis (1995).

In an exemplary embodiment, a sirtuin-modulating compound may traversethe cytoplasmic membrane of a cell. For example, a compound may have acell-permeability of at least about 20%, 50%, 75%, 80%, 90% or 95%.

Sirtuin-modulating compounds described herein may also have one or moreof the following characteristics: the compound may be essentiallynon-toxic to a cell or subject; the sirtuin-modulating compound may bean organic molecule or a small molecule of 2000 amu or less, 1000 amu orless; a compound may have a half-life under normal atmosphericconditions of at least about 30 days, 60 days, 120 days, 6 months or 1year; the compound may have a half-life in solution of at least about 30days, 60 days, 120 days, 6 months or 1 year; a sirtuin-modulatingcompound may be more stable in solution than resveratrol by at least afactor of about 50%, 2 fold, 5 fold, 10 fold, 30 fold, 50 fold or 100fold; a sirtuin-modulating compound may promote deacetylation of the DNArepair factor Ku70; a sirtuin-modulating compound may promotedeacetylation of RelA/p65; a compound may increase general turnoverrates and enhance the sensitivity of cells to TNF-induced apoptosis.

In certain embodiments, a sirtuin-modulating compound does not have anysubstantial ability to inhibit a histone deacetylase (HDACs) class I, aHDAC class II, or HDACs I and II, at concentrations (e.g., in vivo)effective for modulating the deacetylase activity of the sirtuin. Forinstance, in preferred embodiments the sirtuin-modulating compound is asirtuin-activating compound and is chosen to have an EC₅₀ for activatingsirtuin deacetylase activity that is at least 5 fold less than the EC₅₀for inhibition of an HDAC I and/or HDAC II, and even more preferably atleast 10 fold, 100 fold or even 1000 fold less. Methods for assayingHDAC I and/or HDAC II activity are well known in the art and kits toperform such assays may be purchased commercially. See e.g., BioVision,Inc. (Mountain View, Calif.; world wide web at biovision.com) and ThomasScientific (Swedesboro, N.J.; world wide web at tomassci.com).

In certain embodiments, a sirtuin-modulating compound does not have anysubstantial ability to modulate sirtuin homologs. In one embodiment, anactivator of a human sirtuin protein may not have any substantialability to activate a sirtuin protein from lower eukaryotes,particularly yeast or human pathogens, at concentrations (e.g., in vivo)effective for activating the deacetylase activity of human sirtuin. Forexample, a sirtuin-activating compound may be chosen to have an EC₅₀ foractivating a human sirtuin, such as SIRT1 and/or SIRT3, deacetylaseactivity that is at least 5 fold less than the EC₅₀ for activating ayeast sirtuin, such as Sir2 (such as Candida, S. cerevisiae, etc.), andeven more preferably at least 10 fold, 100 fold or even 1000 fold less.In another embodiment, an inhibitor of a sirtuin protein from lowereukaryotes, particularly yeast or human pathogens, does not have anysubstantial ability to inhibit a sirtuin protein from humans atconcentrations (e.g., in vivo) effective for inhibiting the deacetylaseactivity of a sirtuin protein from a lower eukaryote. For example, asirtuin-inhibiting compound may be chosen to have an IC₅₀ for inhibitinga human sirtuin, such as SIRT1 and/or SIRT3, deacetylase activity thatis at least 5 fold less than the IC₅₀ for inhibiting a yeast sirtuin,such as Sir2 (such as Candida, S. cerevisiae, etc.), and even morepreferably at least 10 fold, 100 fold or even 1000 fold less.

In certain embodiments, a sirtuin-modulating compound may have theability to modulate one or more sirtuin protein homologs, such as, forexample, one or more of human SIRT1, SIRT2, SIRT3, SIRT4, SIRT5, SIRT6,or SIRT7. In one embodiment, a sirtuin-modulating compound has theability to modulate both a SIRT1 and a SIRT3 protein.

In other embodiments, a SIRT1 modulator does not have any substantialability to modulate other sirtuin protein homologs, such as, forexample, one or more of human SIRT2, SIRT3, SIRT4, SIRT5, SIRT6, orSIRT7, at concentrations (e.g., in vivo) effective for modulating thedeacetylase activity of human SIRT1. For example, a sirtuin-modulatingcompound may be chosen to have an ED₅₀ for modulating human SIRT1deacetylase activity that is at least 5 fold less than the ED₅₀ formodulating one or more of human SIRT2, SIRT3, SIRT4, SIRT5, SIRT6, orSIRT7, and even more preferably at least 10 fold, 100 fold or even 1000fold less. In one embodiment, a SIRT1 modulator does not have anysubstantial ability to modulate a SIRT3 protein.

In other embodiments, a SIRT3 modulator does not have any substantialability to modulate other sirtuin protein homologs, such as, forexample, one or more of human SIRT1, SIRT2, SIRT4, SIRT5, SIRT6, orSIRT7, at concentrations (e.g., in vivo) effective for modulating thedeacetylase activity of human SIRT3. For example, a sirtuin-modulatingcompound may be chosen to have an ED₅₀ for modulating human SIRT3deacetylase activity that is at least 5 fold less than the ED₅₀ formodulating one or more of human SIRT1, SIRT2, SIRT4, SIRT5, SIRT6, orSIRT7, and even more preferably at least 10 fold, 100 fold or even 1000fold less. In one embodiment, a SIRT3 modulator does not have anysubstantial ability to modulate a SIRT1 protein.

In certain embodiments, a sirtuin-modulating compound may have a bindingaffinity for a sirtuin protein of about 10⁻⁹M, 10⁻¹⁰M, 10⁻¹¹M, 10⁻¹²M orless. A sirtuin-modulating compound may reduce (activator) or increase(inhibitor) the apparent Km of a sirtuin protein for its substrate orNAD+ (or other cofactor) by a factor of at least about 2, 3, 4, 5, 10,20, 30, 50 or 100. In certain embodiments, Km values are determinedusing the mass spectrometry assay described herein. Preferred activatingcompounds reduce the Km of a sirtuin for its substrate or cofactor to agreater extent than caused by resveratrol at a similar concentration orreduce the Km of a sirtuin for its substrate or cofactor similar to thatcaused by resveratrol at a lower concentration. A sirtuin-modulatingcompound may increase the Vmax of a sirtuin protein by a factor of atleast about 2, 3, 4, 5, 10, 20, 30, 50 or 100. A sirtuin-modulatingcompound may have an ED50 for modulating the deacetylase activity of aSIRT1 and/or SIRT3 protein of less than about 1 nM, less than about 10nM, less than about 100 nM, less than about 1 μM, less than about 10 μM,less than about 100 μM, or from about 1-10 nM, from about 10-100 nM,from about 0.1-1 μM, from about 1-10 μM or from about 10-100 μM. Asirtuin-modulating compound may modulate the deacetylase activity of aSIRT1 and/or SIRT3 protein by a factor of at least about 5, 10, 20, 30,50, or 100, as measured in a cellular assay or in a cell based assay. Asirtuin-activating compound may cause at least about 10%, 30%, 50%, 80%,2 fold, 5 fold, 10 fold, 50 fold or 100 fold greater induction of thedeacetylase activity of a sirtuin protein relative to the sameconcentration of resveratrol. A sirtuin-modulating compound may have anED50 for modulating SIRT5 that is at least about 10 fold, 20 fold, 30fold, 50 fold greater than that for modulating SIRT1 and/or SIRT3.

3. Exemplary Uses

In certain aspects, the invention provides methods for modulating thelevel and/or activity of a sirtuin protein and methods of use thereof.

In certain embodiments, the invention provides methods for usingsirtuin-modulating compounds wherein the sirtuin-modulating compoundsactivate a sirtuin protein, e.g., increase the level and/or activity ofa sirtuin protein. Sirtuin-modulating compounds that increase the leveland/or activity of a sirtuin protein may be useful for a variety oftherapeutic applications including, for example, increasing the lifespanof a cell, and treating and/or preventing a wide variety of diseases anddisorders including, for example, diseases or disorders related to agingor stress, diabetes, obesity, neurodegenerative diseases, cardiovasculardisease, blood clotting disorders, inflammation, cancer, and/orflushing, etc. The methods comprise administering to a subject in needthereof a pharmaceutically effective amount of a sirtuin-modulatingcompound, e.g., a sirtuin-activating compound.

In other embodiments, the invention provides methods for usingsirtuin-modulating compounds wherein the sirtuin-modulating compoundsdecrease sirtuin activity, e.g., decrease the level and/or activity of asirtuin protein. Sirtuin-modulating compounds that decrease the leveland/or activity of a sirtuin protein may be useful for a variety oftherapeutic application including, for example, increasing cellularsensitivity to stress (including increasing radiosensitivity and/orchemosensitivity), increasing the amount and/or rate of apoptosis,treatment of cancer (optionally in combination another chemotherapeuticagent), stimulation of appetite, and/or stimulation of weight gain, etc.The methods comprise administering to a subject in need thereof apharmaceutically effective amount of a sirtuin-modulating compound,e.g., a sirtuin-inhibiting compound.

While Applicants do not wish to be bound by theory, it is believed thatactivators and inhibitors of the instant invention may interact with asirtuin at the same location within the sirtuin protein (e.g., activesite or site affecting the Km or Vmax of the active site). It isbelieved that this is the reason why certain classes of sirtuinactivators and inhibitors can have substantial structural similarity.

In certain embodiments, the sirtuin-modulating compounds describedherein may be taken alone or in combination with other compounds. In oneembodiment, a mixture of two or more sirtuin-modulating compounds may beadministered to a subject in need thereof. In another embodiment, asirtuin-modulating compound that increases the level and/or activity ofa sirtuin protein may be administered with one or more of the followingcompounds: resveratrol, butein, fisetin, piceatannol, or quercetin. Inan exemplary embodiment, a sirtuin-modulating compound that increasesthe level and/or activity of a sirtuin protein may be administered incombination with nicotinic acid. In another embodiment, asirtuin-modulating compound that decreases the level and/or activity ofa sirtuin protein may be administered with one or more of the followingcompounds: nicotinamide (NAM), suranim; NF023 (a G-protein antagonist);NF279 (a purinergic receptor antagonist); Trolox(6-hydroxy-2,5,7,8,tetramethylchroman-2-carboxylic acid);(−)-epigallocatechin (hydroxy on sites 3,5,7,3′,4′,5′);(−)-epigallocatechin gallate (Hydroxy sites 5,7,3′,4′,5′ and gallateester on 3); cyanidin choloride (3,5,7,3′,4′-pentahydroxyflavyliumchloride); delphinidin chloride (3,5,7,3′,4′,5′-hexahydroxyflavyliumchloride); myricetin (cannabiscetin; 3,5,7,3′,4′,5′-hexahydroxyflavone);3,7,3′,4′,5′-pentahydroxyflavone; gossypetin(3,5,7,8,3′,4′-hexahydroxyflavone), sirtinol; and splitomicin (see e.g.,Howitz et al. (2003) Nature 425:191; Grozinger et al. (2001) J. Biol.Chem. 276:38837; Dedalov et al. (2001) PNAS 98:15113; and Hirao et al.(2003) J. Biol. Chem 278:52773). In yet another embodiment, one or moresirtuin-modulating compounds may be administered with one or moretherapeutic agents for the treatment or prevention of various diseases,including, for example, cancer, diabetes, neurodegenerative diseases,cardiovascular disease, blood clotting, inflammation, flushing, obesity,ageing, stress, etc. In various embodiments, combination therapiescomprising a sirtuin-modulating compound may refer to (1) pharmaceuticalcompositions that comprise one or more sirtuin-modulating compounds incombination with one or more therapeutic agents (e.g., one or moretherapeutic agents described herein); and (2) co-administration of oneor more sirtuin-modulating compounds with one or more therapeutic agentswherein the sirtuin-modulating compound and therapeutic agent have notbeen formulated in the same compositions (but may be present within thesame kit or package, such as a blister pack or other multi-chamberpackage; connected, separately sealed containers (e.g., foil pouches)that can be separated by the user; or a kit where the sirtuin modulatingcompound(s) and other therapeutic agent(s) are in separate vessels).When using separate formulations, the sirtuin-modulating compound may beadministered at the same, intermittent, staggered, prior to, subsequentto, or combinations thereof, with the administration of anothertherapeutic agent.

In certain embodiments, methods for reducing, preventing or treatingdiseases or disorders using a sirtuin-modulating compound may alsocomprise increasing the protein level of a sirtuin, such as human SIRT1,SIRT2 and/or SIRT3, or homologs thereof. Increasing protein levels canbe achieved by introducing into a cell one or more copies of a nucleicacid that encodes a sirtuin. For example, the level of a sirtuin can beincreased in a mammalian cell by introducing into the mammalian cell anucleic acid encoding the sirtuin, e.g., increasing the level of SIRT1by introducing a nucleic acid encoding the amino acid sequence set forthin GenBank Accession No. NP_(—)036370 and/or increasing the level ofSIRT3 by introducing a nucleic acid encoding the amino acid sequence setforth in GenBank Accession No. AAH01042. The nucleic acid may be underthe control of a promoter that regulates the expression of the SIRT1and/or SIRT3 nucleic acid. Alternatively, the nucleic acid may beintroduced into the cell at a location in the genome that is downstreamof a promoter. Methods for increasing the level of a protein using thesemethods are well known in the art.

A nucleic acid that is introduced into a cell to increase the proteinlevel of a sirtuin may encode a protein that is at least about 80%, 85%,90%, 95%, 98%, or 99% identical to the sequence of a sirtuin, e.g.,SIRT1 (GenBank Accession No. NP_(—)036370) and/or SIRT3 (GenBankAccession No. AAH01042) protein. For example, the nucleic acid encodingthe protein may be at least about 80%, 85%, 90%, 95%, 98%, or 99%identical to a nucleic acid encoding a SIRT1 (e.g. GenBank Accession No.NM_(—)012238) and/or SIRT3 (e.g., GenBank Accession No. BC001042)protein. The nucleic acid may also be a nucleic acid that hybridizes,preferably under stringent hybridization conditions, to a nucleic acidencoding a wild-type sirtuin, e.g., SIRT1 (GenBank Accession No.NM_(—)012238) and/or SIRT3 (e.g., GenBank Accession No. BC001042)protein. Stringent hybridization conditions may include hybridizationand a wash in 0.2×SSC at 65° C. When using a nucleic acid that encodes aprotein that is different from a wild-type sirtuin protein, such as aprotein that is a fragment of a wild-type sirtuin, the protein ispreferably biologically active, e.g., is capable of deacetylation. It isonly necessary to express in a cell a portion of the sirtuin that isbiologically active. For example, a protein that differs from wild-typeSIRT1 having GenBank Accession No. NP_(—)036370, preferably contains thecore structure thereof. The core structure sometimes refers to aminoacids 62-293 of GenBank Accession No. NP_(—)036370, which are encoded bynucleotides 237 to 932 of GenBank Accession No. NM_(—)012238, whichencompasses the NAD binding as well as the substrate binding domains.The core domain of SIRT1 may also refer to about amino acids 261 to 447of GenBank Accession No. NP_(—)036370, which are encoded by nucleotides834 to 1394 of GenBank Accession No. NM_(—)012238; to about amino acids242 to 493 of GenBank Accession No. NP_(—)036370, which are encoded bynucleotides 777 to 1532 of GenBank Accession No. NM_(—)012238; or toabout amino acids 254 to 495 of GenBank Accession No. NP_(—)036370,which are encoded by nucleotides 813 to 1538 of GenBank Accession No.NM_(—)012238. Whether a protein retains a biological function, e.g.,deacetylation capabilities, can be determined according to methods knownin the art.

In certain embodiments, methods for reducing, preventing or treatingdiseases or disorders using a sirtuin-modulating compound may alsocomprise decreasing the protein level of a sirtuin, such as human SIRT1,SIRT2 and/or SIRT3, or homologs thereof. Decreasing a sirtuin proteinlevel can be achieved according to methods known in the art. Forexample, an siRNA, an antisense nucleic acid, or a ribozyme targeted tothe sirtuin can be expressed in the cell. A dominant negative sirtuinmutant, e.g., a mutant that is not capable of deacetylating, may also beused. For example, mutant H363Y of SIRT1, described, e.g., in Luo et al.(2001) Cell 107:137 can be used. Alternatively, agents that inhibittranscription can be used.

Methods for modulating sirtuin protein levels also include methods formodulating the transcription of genes encoding sirtuins, methods forstabilizing/destabilizing the corresponding mRNAs, and other methodsknown in the art.

Aging/Stress

In one embodiment, the invention provides a method extending thelifespan of a cell, extending the proliferative capacity of a cell,slowing ageing of a cell, promoting the survival of a cell, delayingcellular senescence in a cell, mimicking the effects of calorierestriction, increasing the resistance of a cell to stress, orpreventing apoptosis of a cell, by contacting the cell with asirtuin-modulating compound of the invention that increases the leveland/or activity of a sirtuin protein. In an exemplary embodiment, themethods comprise contacting the cell with a sirtuin-activating compound.

The methods described herein may be used to increase the amount of timethat cells, particularly primary cells (i.e., cells obtained from anorganism, e.g., a human), may be kept alive in a cell culture. Embryonicstem (ES) cells and pluripotent cells, and cells differentiatedtherefrom, may also be treated with a sirtuin-modulating compound thatincreases the level and/or activity of a sirtuin protein to keep thecells, or progeny thereof, in culture for longer periods of time. Suchcells can also be used for transplantation into a subject, e.g., afterex vivo modification.

In one embodiment, cells that are intended to be preserved for longperiods of time may be treated with a sirtuin-modulating compound thatincreases the level and/or activity of a sirtuin protein. The cells maybe in suspension (e.g., blood cells, serum, biological growth media,etc.) or in tissues or organs. For example, blood collected from anindividual for purposes of transfusion may be treated with asirtuin-modulating compound that increases the level and/or activity ofa sirtuin protein to preserve the blood cells for longer periods oftime. Additionally, blood to be used for forensic purposes may also bepreserved using a sirtuin-modulating compound that increases the leveland/or activity of a sirtuin protein. Other cells that may be treated toextend their lifespan or protect against apoptosis include cells forconsumption, e.g., cells from non-human mammals (such as meat) or plantcells (such as vegetables).

Sirtuin-modulating compounds that increase the level and/or activity ofa sirtuin protein may also be applied during developmental and growthphases in mammals, plants, insects or microorganisms, in order to, e.g.,alter, retard or accelerate the developmental and/or growth process.

In another embodiment, sirtuin-modulating compounds that increase thelevel and/or activity of a sirtuin protein may be used to treat cellsuseful for transplantation or cell therapy, including, for example,solid tissue grafts, organ transplants, cell suspensions, stem cells,bone marrow cells, etc. The cells or tissue may be an autograft, anallograft, a syngraft or a xenograft. The cells or tissue may be treatedwith the sirtuin-modulating compound prior toadministration/implantation, concurrently withadministration/implantation, and/or post administration/implantationinto a subject. The cells or tissue may be treated prior to removal ofthe cells from the donor individual, ex vivo after removal of the cellsor tissue from the donor individual, or post implantation into therecipient. For example, the donor or recipient individual may be treatedsystemically with a sirtuin-modulating compound or may have a subset ofcells/tissue treated locally with a sirtuin-modulating compound thatincreases the level and/or activity of a sirtuin protein. In certainembodiments, the cells or tissue (or donor/recipient individuals) mayadditionally be treated with another therapeutic agent useful forprolonging graft survival, such as, for example, an immunosuppressiveagent, a cytokine, an angiogenic factor, etc.

In yet other embodiments, cells may be treated with a sirtuin-modulatingcompound that increases the level and/or activity of a sirtuin proteinin vivo, e.g., to increase their lifespan or prevent apoptosis. Forexample, skin can be protected from aging (e.g., developing wrinkles,loss of elasticity, etc.) by treating skin or epithelial cells with asirtuin-modulating compound that increases the level and/or activity ofa sirtuin protein. In an exemplary embodiment, skin is contacted with apharmaceutical or cosmetic composition comprising a sirtuin-modulatingcompound that increases the level and/or activity of a sirtuin protein.Exemplary skin afflictions or skin conditions that may be treated inaccordance with the methods described herein include disorders ordiseases associated with or caused by inflammation, sun damage ornatural aging. For example, the compositions find utility in theprevention or treatment of contact dermatitis (including irritantcontact dermatitis and allergic contact dermatitis), atopic dermatitis(also known as allergic eczema), actinic keratosis, keratinizationdisorders (including eczema), epidermolysis bullosa diseases (includingpenfigus), exfoliative dermatitis, seborrheic dermatitis, erythemas(including erythema multiforme and erythema nodosum), damage caused bythe sun or other light sources, discoid lupus erythematosus,dermatomyositis, psoriasis, skin cancer and the effects of naturalaging. In another embodiment, sirtuin-modulating compounds that increasethe level and/or activity of a sirtuin protein may be used for thetreatment of wounds and/or burns to promote healing, including, forexample, first-, second- or third-degree burns and/or a thermal,chemical or electrical burns. The formulations may be administeredtopically, to the skin or mucosal tissue, as an ointment, lotion, cream,microemulsion, gel, solution or the like, as further described herein,within the context of a dosing regimen effective to bring about thedesired result.

Topical formulations comprising one or more sirtuin-modulating compoundsthat increase the level and/or activity of a sirtuin protein may also beused as preventive, e.g., chemopreventive, compositions. When used in achemopreventive method, susceptible skin is treated prior to any visiblecondition in a particular individual.

Sirtuin-modulating compounds may be delivered locally or systemically toa subject. In one embodiment, a sirtuin-modulating compound is deliveredlocally to a tissue or organ of a subject by injection, topicalformulation, etc.

In another embodiment, a sirtuin-modulating compound that increases thelevel and/or activity of a sirtuin protein may be used for treating orpreventing a disease or condition induced or exacerbated by cellularsenescence in a subject; methods for decreasing the rate of senescenceof a subject, e.g., after onset of senescence; methods for extending thelifespan of a subject; methods for treating or preventing a disease orcondition relating to lifespan; methods for treating or preventing adisease or condition relating to the proliferative capacity of cells;and methods for treating or preventing a disease or condition resultingfrom cell damage or death. In certain embodiments, the method does notact by decreasing the rate of occurrence of diseases that shorten thelifespan of a subject. In certain embodiments, a method does not act byreducing the lethality caused by a disease, such as cancer.

In yet another embodiment, a sirtuin-modulating compound that increasesthe level and/or activity of a sirtuin protein may be administered to asubject in order to generally increase the lifespan of its cells and toprotect its cells against stress and/or against apoptosis. It isbelieved that treating a subject with a compound described herein issimilar to subjecting the subject to hormesis, i.e., mild stress that isbeneficial to organisms and may extend their lifespan.

Sirtuin-modulating compounds that increase the level and/or activity ofa sirtuin protein may be administered to a subject to prevent aging andaging-related consequences or diseases, such as stroke, heart disease,heart failure, arthritis, high blood pressure, and Alzheimer's disease.Other conditions that can be treated include ocular disorders, e.g.,associated with the aging of the eye, such as cataracts, glaucoma, andmacular degeneration. Sirtuin-modulating compounds that increase thelevel and/or activity of a sirtuin protein can also be administered tosubjects for treatment of diseases, e.g., chronic diseases, associatedwith cell death, in order to protect the cells from cell death.Exemplary diseases include those associated with neural cell death,neuronal dysfunction, or muscular cell death or dysfunction, such asParkinson's disease, Alzheimer's disease, multiple sclerosis,amniotropic lateral sclerosis, and muscular dystrophy; AIDS; fulminanthepatitis; diseases linked to degeneration of the brain, such asCreutzfeld-Jakob disease, retinitis pigmentosa and cerebellardegeneration; myelodysplasis such as aplastic anemia; ischemic diseasessuch as myocardial infarction and stroke; hepatic diseases such asalcoholic hepatitis, hepatitis B and hepatitis C; joint-diseases such asosteoarthritis; atherosclerosis; alopecia; damage to the skin due to UVlight; lichen planus; atrophy of the skin; cataract; and graftrejections. Cell death can also be caused by surgery, drug therapy,chemical exposure or radiation exposure.

Sirtuin-modulating compounds that increase the level and/or activity ofa sirtuin protein can also be administered to a subject suffering froman acute disease, e.g., damage to an organ or tissue, e.g., a subjectsuffering from stroke or myocardial infarction or a subject sufferingfrom a spinal cord injury. Sirtuin-modulating compounds that increasethe level and/or activity of a sirtuin protein may also be used torepair an alcoholic's liver.

Cardiovascular Disease

In another embodiment, the invention provides a method for treatingand/or preventing a cardiovascular disease by administering to a subjectin need thereof a sirtuin-modulating compound that increases the leveland/or activity of a sirtuin protein.

Cardiovascular diseases that can be treated or prevented using thesirtuin-modulating compounds that increase the level and/or activity ofa sirtuin protein include cardiomyopathy or myocarditis; such asidiopathic cardiomyopathy, metabolic cardiomyopathy, alcoholiccardiomyopathy, drug-induced cardiomyopathy, ischemic cardiomyopathy,and hypertensive cardiomyopathy. Also treatable or preventable usingcompounds and methods described herein are atheromatous disorders of themajor blood vessels (macrovascular disease) such as the aorta, thecoronary arteries, the carotid arteries, the cerebrovascular arteries,the renal arteries, the iliac arteries, the femoral arteries, and thepopliteal arteries. Other vascular diseases that can be treated orprevented include those related to platelet aggregation, the retinalarterioles, the glomerular arterioles, the vasa nervorum, cardiacarterioles, and associated capillary beds of the eye, the kidney, theheart, and the central and peripheral nervous systems. Thesirtuin-modulating compounds that increase the level and/or activity ofa sirtuin protein may also be used for increasing HDL levels in plasmaof an individual.

Yet other disorders that may be treated with sirtuin-modulatingcompounds that increase the level and/or activity of a sirtuin proteininclude restenosis, e.g., following coronary intervention, and disordersrelating to an abnormal level of high density and low densitycholesterol.

In one embodiment, a sirtuin-modulating compound that increases thelevel and/or activity of a sirtuin protein may be administered as partof a combination therapeutic with another cardiovascular agentincluding, for example, an anti-arrhythmic agent, an antihypertensiveagent, a calcium channel blocker, a cardioplegic solution, a cardiotonicagent, a fibrinolytic agent, a sclerosing solution, a vasoconstrictoragent, a vasodilator agent, a nitric oxide donor, a potassium channelblocker, a sodium channel blocker, statins, or a naturiuretic agent.

In one embodiment, a sirtuin-modulating compound that increases thelevel and/or activity of a sirtuin protein may be administered as partof a combination therapeutic with an anti-arrhythmia agent.Anti-arrhythmia agents are often organized into four main groupsaccording to their mechanism of action: type I, sodium channel blockade;type II, beta-adrenergic blockade; type III, repolarizationprolongation; and type IV, calcium channel blockade. Type Ianti-arrhythmic agents include lidocaine, moricizine, mexiletine,tocamide, procainamide, encamide, flecanide, tocamide, phenyloin,propafenone, quinidine, disopyramide, and flecamide. Type IIanti-arrhythmic agents include propranolol and esmolol. Type IIIincludes agents that act by prolonging the duration of the actionpotential, such as amiodarone, artilide, bretylium, clofilium,isobutilide, sotalol, azimilide, dofetilide, dronedarone, ersentilide,ibutilide, tedisamil, and trecetilide. Type IV anti-arrhythmic agentsinclude verapamil, diltaizem, digitalis, adenosine, nickel chloride, andmagnesium ions.

In another embodiment, a sirtuin-modulating compound that increases thelevel and/or activity of a sirtuin protein may be administered as partof a combination therapeutic with another cardiovascular agent. Examplesof cardiovascular agents include vasodilators, for example, hydralazine;angiotensin converting enzyme inhibitors, for example, captopril;anti-anginal agents, for example, isosorbide nitrate, glyceryltrinitrate and pentaerythritol tetranitrate; anti-arrhythmic agents, forexample, quinidine, procainaltide and lignocaine; cardioglycosides, forexample, digoxin and digitoxin; calcium antagonists, for example,verapamil and nifedipine; diuretics, such as thiazides and relatedcompounds, for example, bendrofluazide, chlorothiazide, chlorothalidone,hydrochlorothiazide and other diuretics, for example, fursemide andtriamterene, and sedatives, for example, nitrazepam, flurazepam anddiazepam.

Other exemplary cardiovascular agents include, for example, acyclooxygenase inhibitor such as aspirin or indomethacin, a plateletaggregation inhibitor such as clopidogrel, ticlopidene or aspirin,fibrinogen antagonists or a diuretic such as chlorothiazide,hydrochlorothiazide, flumethiazide, hydroflumethiazide,bendroflumethiazide, methylchlorthiazide, trichloromethiazide,polythiazide or benzthiazide as well as ethacrynic acid tricrynafen,chlorthalidone, furosemide, musolimine, bumetanide, triamterene,amiloride and spironolactone and salts of such compounds, angiotensinconverting enzyme inhibitors such as captopril, zofenopril, fosinopril,enalapril, ceranopril, cilazopril, delapril, pentopril, quinapril,ramipril, lisinopril, and salts of such compounds, angiotensin IIantagonists such as losartan, irbesartan or valsartan, thrombolyticagents such as tissue plasminogen activator (tPA), recombinant tPA,streptokinase, urokinase, prourokinase, and anisoylated plasminogenstreptokinase activator complex (APSAC, Eminase, Beecham Laboratories),or animal salivary gland plasminogen activators, calcium channelblocking agents such as verapamil, nifedipine or diltiazem, thromboxanereceptor antagonists such as ifetroban, prostacyclin mimetics, orphosphodiesterase inhibitors. Such combination products if formulated asa fixed dose employ the compounds of this invention within the doserange described above and the other pharmaceutically active agent withinits approved dose range.

Yet other exemplary cardiovascular agents include, for example,vasodilators, e.g., bencyclane, cinnarizine, citicoline, cyclandelate,cyclonicate, ebumamonine, phenoxezyl, flunarizine, ibudilast,ifenprodil, lomerizine, naphlole, nikamate, nosergoline, nimodipine,papaverine, pentifylline, nofedoline, vincamin, vinpocetine, vichizyl,pentoxifylline, prostacyclin derivatives (such as prostaglandin E1 andprostaglandin I2), an endothelin receptor blocking drug (such asbosentan), diltiazem, nicorandil, and nitroglycerin. Examples of thecerebral protecting drug include radical scavengers (such as edaravone,vitamin E, and vitamin C), glutamate antagonists, AMPA antagonists,kainate antagonists, NMDA antagonists, GABA agonists, growth factors,opioid antagonists, phosphatidylcholine precursors, serotonin agonists,Na⁺/Ca²⁺ channel inhibitory drugs, and K⁺ channel opening drugs.Examples of the brain metabolic stimulants include amantadine, tiapride,and gamma-aminobutyric acid. Examples of the anticoagulant includeheparins (such as heparin sodium, heparin potassium, dalteparin sodium,dalteparin calcium, heparin calcium, parnaparin sodium, reviparinsodium, and danaparoid sodium), warfarin, enoxaparin, argatroban,batroxobin, and sodium citrate. Examples of the antiplatelet druginclude ticlopidine hydrochloride, dipyridamole, cilostazol, ethylicosapentate, sarpogrelate hydrochloride, dilazep hydrochloride,trapidil, a nonsteroidal antiinflammatory agent (such as aspirin),beraprostsodium, iloprost, and indobufene. Examples of the thrombolyticdrug include urokinase, tissue-type plasminogen activators (such asalteplase, tisokinase, nateplase, pamiteplase, monteplase, andrateplase), and nasaruplase. Examples of the antihypertensive druginclude angiotensin converting enzyme inhibitors (such as captopril,alacepril, lisinopril, imidapril, quinapril, temocapril, delapril,benazepril, cilazapril, trandolapril, enalapril, ceronapril, fosinopril,imadapril, mobertpril, perindopril, ramipril, spirapril, andrandolapril), angiotensin II antagonists (such as losartan, candesartan,valsartan, eprosartan, and irbesartan), calcium channel blocking drugs(such as aranidipine, efonidipine, nicardipine, bamidipine, benidipine,manidipine, cilnidipine, nisoldipine, nitrendipine, nifedipine,nilvadipine, felodipine, amlodipine, diltiazem, bepridil, clentiazem,phendilin, galopamil, mibefradil, prenylamine, semotiadil, terodiline,verapamil, cilnidipine, elgodipine, isradipine, lacidipine,lercanidipine, nimodipine, cinnarizine, flunarizine, lidoflazine,lomerizine, bencyclane, etafenone, and perhexyline), β-adrenalinereceptor blocking drugs (propranolol, pindolol, indenolol, carteolol,bunitrolol, atenolol, acebutolol, metoprolol, timolol, nipradilol,penbutolol, nadolol, tilisolol, carvedilol, bisoprolol, betaxolol,celiprolol, bopindolol, bevantolol, labetalol, alprenolol, amosulalol,arotinolol, befunolol, bucumolol, bufetolol, buferalol, buprandolol,butylidine, butofilolol, carazolol, cetamolol, cloranolol, dilevalol,epanolol, levobunolol, mepindolol, metipranolol, moprolol, nadoxolol,nevibolol, oxprenolol, practol, pronetalol, sotalol, sufinalol,talindolol, tertalol, toliprolol, xybenolol, and esmolol), α-receptorblocking drugs (such as amosulalol, prazosin, terazosin, doxazosin,bunazosin, urapidil, phentolamine, arotinolol, dapiprazole, fenspiride,indoramin, labetalol, naftopidil, nicergoline, tamsulosin, tolazoline,trimazosin, and yohimbine), sympathetic nerve inhibitors (such asclonidine, guanfacine, guanabenz, methyldopa, and reserpine),hydralazine, todralazine, budralazine, and cadralazine. Examples of theantianginal drug include nitrate drugs (such as amyl nitrite,nitroglycerin, and isosorbide), O-adrenaline receptor blocking drugs(such as propranolol, pindolol, indenolol, carteolol, bunitrolol,atenolol, acebutolol, metoprolol, timolol, nipradilol, penbutolol,nadolol, tilisolol, carvedilol, bisoprolol, betaxolol, celiprolol,bopindolol, bevantolol, labetalol, alprenolol, amosulalol, arotinolol,befunolol, bucumolol, bufetolol, buferalol, buprandolol, butylidine,butofilolol, carazolol, cetamolol, cloranolol, dilevalol, epanolol,levobunolol, mepindolol, metipranolol, moprolol, nadoxolol, nevibolol,oxprenolol, practol, pronetalol, sotalol, sufinalol, talindolol,tertalol, toliprolol, andxybenolol), calcium channel blocking drugs(such as aranidipine, efonidipine, nicardipine, bamidipine, benidipine,manidipine, cilnidipine, nisoldipine, nitrendipine, nifedipine,nilvadipine, felodipine, amlodipine, diltiazem, bepridil, clentiazem,phendiline, galopamil, mibefradil, prenylamine, semotiadil, terodiline,verapamil, cilnidipine, elgodipine, isradipine, lacidipine,lercanidipine, nimodipine, cinnarizine, flunarizine, lidoflazine,lomerizine, bencyclane, etafenone, and perhexyline) trimetazidine,dipyridamole, etafenone, dilazep, trapidil, nicorandil, enoxaparin, andaspirin. Examples of the diuretic include thiazide diuretics (such ashydrochlorothiazide, methyclothiazide, trichlormethiazide,benzylhydrochlorothiazide, and penflutizide), loop diuretics (such asfurosemide, etacrynic acid, bumetanide, piretanide, azosemide, andtorasemide), K⁺ sparing diuretics (spironolactone, triamterene,andpotassiumcanrenoate), osmotic diuretics (such as isosorbide,D-mannitol, and glycerin), nonthiazide diuretics (such as meticrane,tripamide, chlorthalidone, and mefruside), and acetazolamide. Examplesof the cardiotonic include digitalis formulations (such as digitoxin,digoxin, methyldigoxin, deslanoside, vesnarinone, lanatoside C, andproscillaridin), xanthine formulations (such as aminophylline, cholinetheophylline, diprophylline, and proxyphylline), catecholamineformulations (such as dopamine, dobutamine, and docarpamine), PDE IIIinhibitors (such as anrinone, olprinone, and milrinone), denopamine,ubidecarenone, pimobendan, levosimendan, aminoethylsulfonic acid,vesnarinone, carperitide, and colforsin daropate. Examples of theantiarrhythmic drug include ajmaline, pirmenol, procainamide,cibenzoline, disopyramide, quinidine, aprindine, mexiletine, lidocaine,phenyloin, pilsicamide, propafenone, flecamide, atenolol, acebutolol,sotalol, propranolol, metoprolol, pindolol, amiodarone, nifekalant,diltiazem, bepridil, and verapamil. Examples of the antihyperlipidemicdrug include atorvastatin, simvastatin, pravastatin sodium, fluvastatinsodium, clinofibrate, clofibrate, simfibrate, fenofibrate, bezafibrate,colestimide, and colestyramine. Examples of the immunosuppressantinclude azathioprine, mizoribine, cyclosporine, tacrolimus, gusperimus,and methotrexate.

Cell Death/Cancer

Sirtuin-modulating compounds that increase the level and/or activity ofa sirtuin protein may be administered to subjects who have recentlyreceived or are likely to receive a dose of radiation or toxin. In oneembodiment, the dose of radiation or toxin is received as part of awork-related or medical procedure, e.g., working in a nuclear powerplant, flying an airplane, an X-ray, CAT scan, or the administration ofa radioactive dye for medical imaging; in such an embodiment, thecompound is administered as a prophylactic measure. In anotherembodiment, the radiation or toxin exposure is received unintentionally,e.g., as a result of an industrial accident, habitation in a location ofnatural radiation, terrorist act, or act of war involving radioactive ortoxic material. In such a case, the compound is preferably administeredas soon as possible after the exposure to inhibit apoptosis and thesubsequent development of acute radiation syndrome.

Sirtuin-modulating compounds may also be used for treating and/orpreventing cancer. In certain embodiments, sirtuin-modulating compoundsthat increase the level and/or activity of a sirtuin protein may be usedfor treating and/or preventing cancer. Calorie restriction has beenlinked to a reduction in the incidence of age-related disordersincluding cancer (see e.g., Bordone and Guarente, Nat. Rev. Mol. CellBiol. (2005 epub); Guarente and Picard, Cell 120: 473-82 (2005);Berrigan, et al., Carcinogenesis 23: 817-822 (2002); and Heilbronn andRavussin, Am. J. Clin. Nutr. 78: 361-369 (2003)). Additionally, the Sir2protein from yeast has been shown to be required for lifespan extensionby glucose restriction (see e.g., Lin et al., Science 289: 2126-2128(2000); Anderson et al., Nature 423: 181-185 (2003)), a yeast model forcalorie restriction. Accordingly, an increase in the level and/oractivity of a sirtuin protein may be useful for treating and/orpreventing the incidence of age-related disorders, such as, for example,cancer. In other embodiments, sirtuin-modulating compounds that decreasethe level and/or activity of a sirtuin protein may be used for treatingor preventing cancer. For example, inhibitory compounds may be used tostimulate acetylation of substrates such as p53 and thereby increaseapoptosis, as well as to reduce the lifespan of cells and organisms,render them more sensitive to stress, and/or increase theradiosensitivity and/or chemosensitivity of a cell or organism. Thus,inhibitory compounds may be used, e.g., for treating cancer. Exemplarycancers that may be treated using a sirtuin-modulating compound arethose of the brain and kidney; hormone-dependent cancers includingbreast, prostate, testicular, and ovarian cancers; lymphomas, andleukemias. In cancers associated with solid tumors, a modulatingcompound may be administered directly into the tumor. Cancer of bloodcells, e.g., leukemia, can be treated by administering a modulatingcompound into the blood stream or into the bone marrow. Benign cellgrowth can also be treated, e.g., warts. Other diseases that can betreated include autoimmune diseases, e.g., systemic lupus erythematosus,scleroderma, and arthritis, in which autoimmune cells should be removed.Viral infections such as herpes, HIV, adenovirus, and HTLV-1 associatedmalignant and benign disorders can also be treated by administration ofsirtuin-modulating compound. Alternatively, cells can be obtained from asubject, treated ex vivo to remove certain undesirable cells, e.g.,cancer cells, and administered back to the same or a different subject.

Chemotherapeutic agents that may be coadministered with modulatingcompounds described herein as having anti-cancer activity (e.g.,compounds that induce apoptosis, compounds that reduce lifespan orcompounds that render cells sensitive to stress) include:aminoglutethimide, amsacrine, anastrozole, asparaginase, bcg,bicalutamide, bleomycin, buserelin, busulfan, campothecin, capecitabine,carboplatin, carmustine, chlorambucil, cisplatin, cladribine,clodronate, colchicine, cyclophosphamide, cyproterone, cytarabine,dacarbazine, dactinomycin, daunorubicin, dienestrol, diethylstilbestrol,docetaxel, doxorubicin, epirubicin, estradiol, estramustine, etoposide,exemestane, filgrastim, fludarabine, fludrocortisone, fluorouracil,fluoxymesterone, flutamide, gemcitabine, genistein, goserelin,hydroxyurea, idarubicin, ifosfamide, imatinib, interferon, irinotecan,ironotecan, letrozole, leucovorin, leuprolide, levamisole, lomustine,mechlorethamine, medroxyprogesterone, megestrol, melphalan,mercaptopurine, mesna, methotrexate, mitomycin, mitotane, mitoxantrone,nilutamide, nocodazole, octreotide, oxaliplatin, paclitaxel,pamidronate, pentostatin, plicamycin, porfimer, procarbazine,raltitrexed, rituximab, streptozocin, suramin, tamoxifen, temozolomide,teniposide, testosterone, thioguanine, thiotepa, titanocene dichloride,topotecan, trastuzumab, tretinoin, vinblastine, vincristine, vindesine,and vinorelbine.

These chemotherapeutic agents may be categorized by their mechanism ofaction into, for example, following groups: anti-metabolites/anti-canceragents, such as pyrimidine analogs (5-fluorouracil, floxuridine,capecitabine, gemcitabine and cytarabine) and purine analogs, folateantagonists and related inhibitors (mercaptopurine, thioguanine,pentostatin and 2-chlorodeoxyadenosine (cladribine));antiproliferative/antimitotic agents including natural products such asvinca alkaloids (vinblastine, vincristine, and vinorelbine), microtubuledisruptors such as taxane (paclitaxel, docetaxel), vincristin,vinblastin, nocodazole, epothilones and navelbine, epidipodophyllotoxins(teniposide), DNA damaging agents (actinomycin, amsacrine,anthracyclines, bleomycin, busulfan, camptothecin, carboplatin,chlorambucil, cisplatin, cyclophosphamide, cytoxan, dactinomycin,daunorubicin, docetaxel, doxorubicin, epirubicin,hexamethylmelamineoxaliplatin, iphosphamide, melphalan,merchlorethamine, mitomycin, mitoxantrone, nitrosourea, paclitaxel,plicamycin, procarbazine, teniposide, triethylenethiophosphoramide andetoposide (VP16)); antibiotics such as dactinomycin (actinomycin D),daunorubicin, doxorubicin (adriamycin), idarubicin, anthracyclines,mitoxantrone, bleomycins, plicamycin (mithramycin) and mitomycin;enzymes (L-asparaginase which systemically metabolizes L-asparagine anddeprives cells which do not have the capacity to synthesize their ownasparagine); antiplatelet agents; antiproliferative/antimitoticalkylating agents such as nitrogen mustards (mechlorethamine,cyclophosphamide and analogs, melphalan, chlorambucil), ethyleniminesand methylmelamines (hexamethylmelamine and thiotepa), alkylsulfonates-busulfan, nitrosoureas (carmustine (BCNU) and analogs,streptozocin), trazenes—dacarbazinine (DTIC);antiproliferative/antimitotic antimetabolites such as folic acid analogs(methotrexate); platinum coordination complexes (cisplatin,carboplatin), procarbazine, hydroxyurea, mitotane, aminoglutethimide;hormones, hormone analogs (estrogen, tamoxifen, goserelin, bicalutamide,nilutamide) and aromatase inhibitors (letrozole, anastrozole);anticoagulants (heparin, synthetic heparin salts and other inhibitors ofthrombin); fibrinolytic agents (such as tissue plasminogen activator,streptokinase and urokinase), aspirin, COX-2 inhibitors, dipyridamole,ticlopidine, clopidogrel, abciximab; antimigratory agents; antisecretoryagents (breveldin); immunosuppressives (cyclosporine, tacrolimus(FK-506), sirolimus (rapamycin), azathioprine, mycophenolate mofetil);anti-angiogenic compounds (TNP470, genistein) and growth factorinhibitors (vascular endothelial growth factor (VEGF) inhibitors,fibroblast growth factor (FGF) inhibitors, epidermal growth factor (EGF)inhibitors); angiotensin receptor blocker; nitric oxide donors;anti-sense oligonucleotides; antibodies (trastuzumab); cell cycleinhibitors and differentiation inducers (tretinoin); mTOR inhibitors,topoisomerase inhibitors (doxorubicin (adriamycin), amsacrine,camptothecin, daunorubicin, dactinomycin, eniposide, epirubicin,etoposide, idarubicin, irinotecan (CPT-11) and mitoxantrone, topotecan,irinotecan), corticosteroids (cortisone, dexamethasone, hydrocortisone,methylpednisolone, prednisone, and prenisolone); growth factor signaltransduction kinase inhibitors; mitochondrial dysfunction inducers andcaspase activators; chromatin disruptors.

These chemotherapeutic agents may be used by themselves with asirtuin-modulating compound described herein as inducing cell death orreducing lifespan or increasing sensitivity to stress and/or incombination with other chemotherapeutics agents. Many combinatorialtherapies have been developed, including but not limited to those listedin Table 1.

TABLE 1 Exemplary combinatorial therapies for the treatment of cancer.Name Therapeutic agents ABV Doxorubicin, Bleomycin, Vinblastine ABVDDoxorubicin, Bleomycin, Vinblastine, Dacarbazine AC (Breast)Doxorubicin, Cyclophosphamide AC (Sarcoma) Doxorubicin, Cisplatin AC(Neuroblastoma) Cyclophosphamide, Doxorubicin ACE Cyclophosphamide,Doxorubicin, Etoposide ACe Cyclophosphamide, Doxorubicin AD Doxorubicin,Dacarbazine AP Doxorubicin, Cisplatin ARAC-DNR Cytarabine, DaunorubicinB-CAVe Bleomycin, Lomustine, Doxorubicin, Vinblastine BCVPP Carmustine,Cyclophosphamide, Vinblastine, Procarbazine, Prednisone BEACOPPBleomycin, Etoposide, Doxorubicin, Cyclophosphamide, Vincristine,Procarbazine, Prednisone, Filgrastim BEP Bleomycin, Etoposide, CisplatinBIP Bleomycin, Cisplatin, Ifosfamide, Mesna BOMP Bleomycin, Vincristine,Cisplatin, Mitomycin CA Cytarabine, Asparaginase CABO Cisplatin,Methotrexate, Bleomycin, Vincristine CAF Cyclophosphamide, Doxorubicin,Fluorouracil CAL-G Cyclophosphamide, Daunorubicin, Vincristine,Prednisone, Asparaginase CAMP Cyclophosphamide, Doxorubicin,Methotrexate, Procarbazine CAP Cyclophosphamide, Doxorubicin, CisplatinCaT Carboplatin, Paclitaxel CAV Cyclophosphamide, Doxorubicin,Vincristine CAVE ADD CAV and Etoposide CA-VP16 Cyclophosphamide,Doxorubicin, Etoposide CC Cyclophosphamide, Carboplatin CDDP/VP-16Cisplatin, Etoposide CEF Cyclophosphamide, Epirubicin, FluorouracilCEPP(B) Cyclophosphamide, Etoposide, Prednisone, with or without/Bleomycin CEV Cyclophosphamide, Etoposide, Vincristine CF Cisplatin,Fluorouracil or Carboplatin Fluorouracil CHAP Cyclophosphamide orCyclophosphamide, Altretamine, Doxorubicin, Cisplatin ChlVPPChlorambucil, Vinblastine, Procarbazine, Prednisone CHOPCyclophosphamide, Doxorubicin, Vincristine, Prednisone CHOP-BLEO AddBleomycin to CHOP CISCA Cyclophosphamide, Doxorubicin, CisplatinCLD-BOMP Bleomycin, Cisplatin, Vincristine, Mitomycin CMF Methotrexate,Fluorouracil, Cyclophosphamide CMFP Cyclophosphamide, Methotrexate,Fluorouracil, Prednisone CMFVP Cyclophosphamide, Methotrexate,Fluorouracil, Vincristine, Prednisone CMV Cisplatin, Methotrexate,Vinblastine CNF Cyclophosphamide, Mitoxantrone, Fluorouracil CNOPCyclophosphamide, Mitoxantrone, Vincristine, Prednisone COB Cisplatin,Vincristine, Bleomycin CODE Cisplatin, Vincristine, Doxorubicin,Etoposide COMLA Cyclophosphamide, Vincristine, Methotrexate, Leucovorin,Cytarabine COMP Cyclophosphamide, Vincristine, Methotrexate, PrednisoneCooper Regimen Cyclophosphamide, Methotrexate, Fluorouracil,Vincristine, Prednisone COP Cyclophosphamide, Vincristine, PrednisoneCOPE Cyclophosphamide, Vincristine, Cisplatin, Etoposide COPPCyclophosphamide, Vincristine, Procarbazine, Prednisone CP(Chroniclymphocytic Chlorambucil, Prednisone leukemia) CP (Ovarian Cancer)Cyclophosphamide, Cisplatin CT Cisplatin, Paclitaxel CVD Cisplatin,Vinblastine, Dacarbazine CVI Carboplatin, Etoposide, Ifosfamide, MesnaCVP Cyclophosphamide, Vincristine, Prednisome CVPP Lomustine,Procarbazine, Prednisone CYVADIC Cyclophosphamide, Vincristine,Doxorubicin, Dacarbazine DA Daunorubicin, Cytarabine DAT Daunorubicin,Cytarabine, Thioguanine DAV Daunorubicin, Cytarabine, Etoposide DCTDaunorubicin, Cytarabine, Thioguanine DHAP Cisplatin, Cytarabine,Dexamethasone DI Doxorubicin, Ifosfamide DTIC/Tamoxifen Dacarbazine,Tamoxifen DVP Daunorubicin, Vincristine, Prednisone EAP Etoposide,Doxorubicin, Cisplatin EC Etoposide, Carboplatin EFP Etoposie,Fluorouracil, Cisplatin ELF Etoposide, Leucovorin, Fluorouracil EMA 86Mitoxantrone, Etoposide, Cytarabine EP Etoposide, Cisplatin EVAEtoposide, Vinblastine FAC Fluorouracil, Doxorubicin, CyclophosphamideFAM Fluorouracil, Doxorubicin, Mitomycin FAMTX Methotrexate, Leucovorin,Doxorubicin FAP Fluorouracil, Doxorubicin, Cisplatin F-CL Fluorouracil,Leucovorin FEC Fluorouracil, Cyclophosphamide, Epirubicin FEDFluorouracil, Etoposide, Cisplatin FL Flutamide, Leuprolide FZFlutamide, Goserelin acetate implant HDMTX Methotrexate, LeucovorinHexa-CAF Altretamine, Cyclophosphamide, Methotrexate, Fluorouracil ICE-TIfosfamide, Carboplatin, Etoposide, Paclitaxel, Mesna IDMTX/6-MPMethotrexate, Mercaptopurine, Leucovorin IE Ifosfamide, Etoposie, MesnaIfoVP Ifosfamide, Etoposide, Mesna IPA Ifosfamide, Cisplatin,Doxorubicin M-2 Vincristine, Carmustine, Cyclophosphamide, Prednisone,Melphalan MAC-III Methotrexate, Leucovorin, Dactinomycin,Cyclophosphamide MACC Methotrexate, Doxorubicin, Cyclophosphamide,Lomustine MACOP-B Methotrexate, Leucovorin, Doxorubicin,Cyclophosphamide, Vincristine, Bleomycin, Prednisone MAID Mesna,Doxorubicin, Ifosfamide, Dacarbazine m-BACOD Bleomycin, Doxorubicin,Cyclophosphamide, Vincristine, Dexamethasone, Methotrexate, LeucovorinMBC Methotrexate, Bleomycin, Cisplatin MC Mitoxantrone, Cytarabine MFMethotrexate, Fluorouracil, Leucovorin MICE Ifosfamide, Carboplatin,Etoposide, Mesna MINE Mesna, Ifosfamide, Mitoxantrone, Etoposidemini-BEAM Carmustine, Etoposide, Cytarabine, Melphalan MOBP Bleomycin,Vincristine, Cisplatin, Mitomycin MOP Mechlorethamine, Vincristine,Procarbazine MOPP Mechlorethamine, Vincristine, Procarbazine, PrednisoneMOPP/ABV Mechlorethamine, Vincristine, Procarbazine, Prednisone,Doxorubicin, Bleomycin, Vinblastine MP (multiple myeloma) Melphalan,Prednisone MP (prostate cancer) Mitoxantrone, Prednisone MTX/6-MOMethotrexate, Mercaptopurine MTX/6-MP/VP Methotrexate, Mercaptopurine,Vincristine, Prednisone MTX-CDDPAdr Methotrexate, Leucovorin, Cisplatin,Doxorubicin MV (breast cancer) Mitomycin, Vinblastine MV (acutemyelocytic Mitoxantrone, Etoposide leukemia) M-VAC MethotrexateVinblastine, Doxorubicin, Cisplatin MVP Mitomycin Vinblastine, CisplatinMVPP Mechlorethamine, Vinblastine, Procarbazine, Prednisone NFLMitoxantrone, Fluorouracil, Leucovorin NOVP Mitoxantrone, Vinblastine,Vincristine OPA Vincristine, Prednisone, Doxorubicin OPPA AddProcarbazine to OPA. PAC Cisplatin, Doxorubicin PAC-I Cisplatin,Doxorubicin, Cyclophosphamide PA-CI Cisplatin, Doxorubicin PCPaclitaxel, Carboplatin or Paclitaxel, Cisplatin PCV Lomustine,Procarbazine, Vincristine PE Paclitaxel, Estramustine PFL Cisplatin,Fluorouracil, Leucovorin POC Prednisone, Vincristine, Lomustine ProMACEPrednisone, Methotrexate, Leucovorin, Doxorubicin, Cyclophosphamide,Etoposide ProMACE/cytaBOM Prednisone, Doxorubicin, Cyclophosphamide,Etoposide, Cytarabine, Bleomycin, Vincristine, Methotrexate, Leucovorin,Cotrimoxazole PRoMACE/MOPP Prednisone, Doxorubicin, Cyclophosphamide,Etoposide, Mechlorethamine, Vincristine, Procarbazine, Methotrexate,Leucovorin Pt/VM Cisplatin, Teniposide PVA Prednisone, Vincristine,Asparaginase PVB Cisplatin, Vinblastine, Bleomycin PVDA Prednisone,Vincristine, Daunorubicin, Asparaginase SMF Streptozocin, Mitomycin,Fluorouracil TAD Mechlorethamine, Doxorubicin, Vinblastine, Vincristine,Bleomycin, Etoposide, Prednisone TCF Paclitaxel, Cisplatin, FluorouracilTIP Paclitaxel, Ifosfamide, Mesna, Cisplatin TTT Methotrexate,Cytarabine, Hydrocortisone Topo/CTX Cyclophosphamide, Topotecan, MesnaVAB-6 Cyclophosphamide, Dactinomycin, Vinblastine, Cisplatin, BleomycinVAC Vincristine, Dactinomycin, Cyclophosphamide VACAdr Vincristine,Cyclophosphamide, Doxorubicin, Dactinomycin, Vincristine VADVincristine, Doxorubicin, Dexamethasone VATH Vinblastine, Doxorubicin,Thiotepa, Flouxymesterone VBAP Vincristine, Carmustine, Doxorubicin,Prednisone VBCMP Vincristine, Carmustine, Melphalan, Cyclophosphamide,Prednisone VC Vinorelbine, Cisplatin VCAP Vincristine, Cyclophosphamide,Doxorubicin, Prednisone VD Vinorelbine, Doxorubicin VelP Vinblastine,Cisplatin, Ifosfamide, Mesna VIP Etoposide, Cisplatin, Ifosfamide, MesnaVM Mitomycin, Vinblastine VMCP Vincristine, Melphalan, Cyclophosphamide,Prednisone VP Etoposide, Cisplatin V-TAD Etoposide, Thioguanine,Daunorubicin, Cytarabine 5 + 2 Cytarabine, Daunorubicin, Mitoxantrone7 + 3 Cytarabine with/, Daunorubicin or Idarubicin or Mitoxantrone “8 in1” Methylprednisolone, Vincristine, Lomustine, Procarbazine,Hydroxyurea, Cisplatin, Cytarabine, Dacarbazine

In addition to conventional chemotherapeutics, the sirtuin-modulatingcompounds described herein as capable of inducing cell death or reducinglifespan can also be used with antisense RNA, RNAi or otherpolynucleotides to inhibit the expression of the cellular componentsthat contribute to unwanted cellular proliferation that are targets ofconventional chemotherapy. Such targets are, merely to illustrate,growth factors, growth factor receptors, cell cycle regulatory proteins,transcription factors, or signal transduction kinases.

Combination therapies comprising sirtuin-modulating compounds and aconventional chemotherapeutic agent may be advantageous over combinationtherapies known in the art because the combination allows theconventional chemotherapeutic agent to exert greater effect at lowerdosage. In a preferred embodiment, the effective dose (ED₅₀) for achemotherapeutic agent, or combination of conventional chemotherapeuticagents, when used in combination with a sirtuin-modulating compound isat least 2 fold less than the ED₅₀ for the chemotherapeutic agent alone,and even more preferably at 5 fold, 10 fold or even 25 fold less.Conversely, the therapeutic index (TI) for such chemotherapeutic agentor combination of such chemotherapeutic agent when used in combinationwith a sirtuin-modulating compound described herein can be at least 2fold greater than the TI for conventional chemotherapeutic regimenalone, and even more preferably at 5 fold, 10 fold or even 25 foldgreater.

Neuronal Diseases/Disorders

In certain aspects, sirtuin-modulating compounds that increase the leveland/or activity of a sirtuin protein can be used to treat patientssuffering from neurodegenerative diseases, and traumatic or mechanicalinjury to the central nervous system (CNS), spinal cord or peripheralnervous system (PNS). Neurodegenerative disease typically involvesreductions in the mass and volume of the human brain, which may be dueto the atrophy and/or death of brain cells, which are far more profoundthan those in a healthy person that are attributable to aging.Neurodegenerative diseases can evolve gradually, after a long period ofnormal brain function, due to progressive degeneration (e.g., nerve celldysfunction and death) of specific brain regions. Alternatively,neurodegenerative diseases can have a quick onset, such as thoseassociated with trauma or toxins. The actual onset of brain degenerationmay precede clinical expression by many years. Examples ofneurodegenerative diseases include, but are not limited to, Alzheimer'sdisease (AD), Parkinson's disease (PD), Huntington's disease (HD),amyotrophic lateral sclerosis (ALS; Lou Gehrig's disease), diffuse Lewybody disease, chorea-acanthocytosis, primary lateral sclerosis, oculardiseases (ocular neuritis), chemotherapy-induced neuropathies (e.g.,from vincristine, paclitaxel, bortezomib), diabetes-induced neuropathiesand Friedreich's ataxia. Sirtuin-modulating compounds that increase thelevel and/or activity of a sirtuin protein can be used to treat thesedisorders and others as described below.

AD is a chronic, incurable, and unstoppable CNS disorder that occursgradually, resulting in memory loss, unusual behavior, personalitychanges, and a decline in thinking abilities. These losses are relatedto the death of specific types of brain cells and the breakdown ofconnections and their supporting network (e.g. glial cells) betweenthem. AD has been described as childhood development in reverse. In mostpeople with AD, symptoms appear after the age 60. The earliest symptomsinclude loss of recent memory, faulty judgment, and changes inpersonality. Later in the disease, those with AD may forget how to dosimple tasks like washing their hands. Eventually people with AD loseall reasoning abilities and become dependent on other people for theireveryday care. Finally, the disease becomes so debilitating thatpatients are bedridden and typically develop coexisting illnesses.

PD is a chronic, incurable, and unstoppable CNS disorder that occursgradually and results in uncontrolled body movements, rigidity, tremor,and dyskinesia. These motor system problems are related to the death ofbrain cells in an area of the brain that produces dopamine, a chemicalthat helps control muscle activity. In most people with PD, symptomsappear after age 50. The initial symptoms of PD are a pronounced tremoraffecting the extremities, notably in the hands or lips. Subsequentcharacteristic symptoms of PD are stiffness or slowness of movement, ashuffling walk, stooped posture, and impaired balance. There are wideranging secondary symptoms such as memory loss, dementia, depression,emotional changes, swallowing difficulties, abnormal speech, sexualdysfunction, and bladder and bowel problems. These symptoms will beginto interfere with routine activities, such as holding a fork or readinga newspaper. Finally, people with PD become so profoundly disabled thatthey are bedridden.

ALS (motor neuron disease) is a chronic, incurable, and unstoppable CNSdisorder that attacks the motor neurons, components of the CNS thatconnect the brain to the skeletal muscles. In ALS, the motor neuronsdeteriorate and eventually die, and though a person's brain normallyremains fully functioning and alert, the command to move never reachesthe muscles. Most people who get ALS are between 40 and 70 years old.The first motor neurons that weaken are those controlling the arms orlegs. Those with ALS may have trouble walking, they may drop things,fall, slur their speech, and laugh or cry uncontrollably. Eventually themuscles in the limbs begin to atrophy from disuse. This muscle weaknesswill become debilitating and a person will need a wheel chair or becomeunable to function out of bed.

The causes of these neurological diseases have remained largely unknown.They are conventionally defined as distinct diseases, yet clearly showextraordinary similarities in basic processes and commonly demonstrateoverlapping symptoms far greater than would be expected by chance alone.Current disease definitions fail to properly deal with the issue ofoverlap and a new classification of the neurodegenerative disorders hasbeen called for.

HD is another neurodegenerative disease resulting from geneticallyprogrammed degeneration of neurons in certain areas of the brain. Thisdegeneration causes uncontrolled movements, loss of intellectualfaculties, and emotional disturbance. HD is a familial disease, passedfrom parent to child through a dominant mutation in the wild-type gene.Some early symptoms of HD are mood swings, depression, irritability ortrouble driving, learning new things, remembering a fact, or making adecision. As the disease progresses, concentration on intellectual tasksbecomes increasingly difficult and the patient may have difficultyfeeding himself or herself and swallowing.

Tay-Sachs disease and Sandhoff disease are glycolipid storage diseasescaused by the lack of lysosomal β-hexosaminidase (Gravel et al., in TheMetabolic Basis of Inherited Disease, eds. Scriver et al., McGraw-Hill,New York, pp. 2839-2879, 1995). In both disorders, GM2 ganglioside andrelated glycolipidssubstrates for β-hexosaminidase accumulate in thenervous system and trigger acute neurodegeneration. In the most severeforms, the onset of symptoms begins in early infancy. A precipitousneurodegenerative course then ensues, with affected infants exhibitingmotor dysfunction, seizure, visual loss, and deafness. Death usuallyoccurs by 2-5 years of age. Neuronal loss through an apoptotic mechanismhas been demonstrated (Huang et al., Hum. Mol. Genet. 6: 1879-1885,1997).

It is well-known that apoptosis plays a role in AIDS pathogenesis in theimmune system. However, HIV-1 also induces neurological disease. Shi etal. (J. Clin. Invest. 98: 1979-1990, 1996) examined apoptosis induced byHIV-1 infection of the CNS in an in vitro model and in brain tissue fromAIDS patients, and found that HIV-1 infection of primary brain culturesinduced apoptosis in neurons and astrocytes in vitro. Apoptosis ofneurons and astrocytes was also detected in brain tissue from 10/11 AIDSpatients, including 5/5 patients with HIV-1 dementia and 4/5 nondementedpatients.

There are four main peripheral neuropathies associated with HIV, namelysensory neuropathy, AIDP/CIPD, drug-induced neuropathy and CMV-related.

The most common type of neuropathy associated with AIDS is distalsymmetrical polyneuropathy (DSPN). This syndrome is a result of nervedegeneration and is characterized by numbness and a sensation of pinsand needles. DSPN causes few serious abnormalities and mostly results innumbness or tingling of the feet and slowed reflexes at the ankles. Itgenerally occurs with more severe immunosuppression and is steadilyprogressive. Treatment with tricyclic antidepressants relieves symptomsbut does not affect the underlying nerve damage.

A less frequent, but more severe type of neuropathy is known as acute orchronic inflammatory demyelinating polyneuropathy (AIDP/CIDP). InAIDP/CIDP there is damage to the fatty membrane covering the nerveimpulses. This kind of neuropathy involves inflammation and resemblesthe muscle deterioration often identified with long-term use of AZT. Itcan be the first manifestation of HIV infection, where the patient maynot complain of pain, but fails to respond to standard reflex tests.This kind of neuropathy may be associated with seroconversion, in whichcase it can sometimes resolve spontaneously. It can serve as a sign ofHIV infection and indicate that it might be time to consider antiviraltherapy. AIDP/CIDP may be auto-immune in origin.

Drug-induced, or toxic, neuropathies can be very painful. Antiviraldrugs commonly cause peripheral neuropathy, as do other drugs e.g.vincristine, dilantin (an anti-seizure medication), high-dose vitamins,isoniazid, and folic acid antagonists. Peripheral neuropathy is oftenused in clinical trials for antivirals as a dose-limiting side effect,which means that more drugs should not be administered. Additionally,the use of such drugs can exacerbate otherwise minor neuropathies.Usually, these drug-induced neuropathies are reversible with thediscontinuation of the drug.

CMV causes several neurological syndromes in AIDS, includingencephalitis, myelitis, and polyradiculopathy.

Neuronal loss is also a salient feature of prion diseases, such asCreutzfeldt-Jakob disease in human, BSE in cattle (mad cow disease),Scrapie Disease in sheep and goats, and feline spongiform encephalopathy(FSE) in cats. Sirtuin-modulating compounds that increase the leveland/or activity of a sirtuin protein may be useful for treating orpreventing neuronal loss due to these prior diseases.

In another embodiment, a sirtuin-modulating compound that increases thelevel and/or activity of a sirtuin protein may be used to treat orprevent any disease or disorder involving axonopathy. Distal axonopathyis a type of peripheral neuropathy that results from some metabolic ortoxic derangement of peripheral nervous system (PNS) neurons. It is themost common response of nerves to metabolic or toxic disturbances, andas such may be caused by metabolic diseases such as diabetes, renalfailure, deficiency syndromes such as malnutrition and alcoholism, orthe effects of toxins or drugs. The most common cause of distalaxonopathy is diabetes, and the most common distal axonopathy isdiabetic neuropathy. The most distal portions of axons are usually thefirst to degenerate, and axonal atrophy advances slowly towards thenerve's cell body. If the noxious stimulus is removed, regeneration ispossible, though prognosis decreases depending on the duration andseverity of the stimulus. Those with distal axonopathies usually presentwith symmetrical glove-stocking sensori-motor disturbances. Deep tendonreflexes and autonomic nervous system (ANS) functions are also lost ordiminished in affected areas.

Diabetic neuropathies are neuropathic disorders that are associated withdiabetes mellitus. These conditions usually result from diabeticmicrovascular injury involving small blood vessels that supply nerves(vasa nervorum). Relatively common conditions which may be associatedwith diabetic neuropathy include third nerve palsy; mononeuropathy;mononeuritis multiplex; diabetic amyotrophy; a painful polyneuropathy;autonomic neuropathy; and thoracoabdominal neuropathy. Clinicalmanifestations of diabetic neuropathy include, for example, sensorimotorpolyneuropathy such as numbness, sensory loss, dysesthesia and nighttimepain; autonomic neuropathy such as delayed gastric emptying orgastroparesis; and cranial neuropathy such as oculomotor (3rd)neuropathies or Mononeuropathies of the thoracic or lumbar spinalnerves.

Peripheral neuropathy is the medical term for damage to nerves of theperipheral nervous system, which may be caused either by diseases of thenerve or from the side-effects of systemic illness. Peripheralneuropathies vary in their presentation and origin, and may affect thenerve or the neuromuscular junction. Major causes of peripheralneuropathy include seizures, nutritional deficiencies, and HIV, thoughdiabetes is the most likely cause. Mechanical pressure from staying inone position for too long, a tumor, intraneural hemorrhage, exposing thebody to extreme conditions such as radiation, cold temperatures, ortoxic substances can also cause peripheral neuropathy.

In an exemplary embodiment, a sirtuin-modulating compound that increasesthe level and/or activity of a sirtuin protein may be used to treat orprevent multiple sclerosis (MS), including relapsing MS andmonosymptomatic MS, and other demyelinating conditions, such as, forexample, chromic inflammatory demyelinating polyneuropathy (CIDP), orsymptoms associated therewith.

MS is a chronic, often disabling disease of the central nervous system.Various and converging lines of evidence point to the possibility thatthe disease is caused by a disturbance in the immune function, althoughthe cause of this disturbance has not been established. This disturbancepermits cells of the immune system to “attack” myelin, the fatcontaining insulating sheath that surrounds the nerve axons located inthe central nervous system (“CNS”). When myelin is damaged, electricalpulses cannot travel quickly or normally along nerve fiber pathways inthe brain and spinal cord. This results in disruption of normalelectrical conductivity within the axons, fatigue and disturbances ofvision, strength, coordination, balance, sensation, and bladder andbowel function.

As such, MS is now a common and well-known neurological disorder that ischaracterized by episodic patches of inflammation and demyelinationwhich can occur anywhere in the CNS. However, almost always without anyinvolvement of the peripheral nerves associated therewith. Demyelinationproduces a situation analogous to that resulting from cracks or tears inan insulator surrounding an electrical cord. That is, when theinsulating sheath is disrupted, the circuit is “short circuited” and theelectrical apparatus associated therewith will function intermittentlyor nor at all. Such loss of myelin surrounding nerve fibers results inshort circuits in nerves traversing the brain and the spinal cord thatthereby result in symptoms of MS. It is further found that suchdemyelination occurs in patches, as opposed to along the entire CNS. Inaddition, such demyelination may be intermittent. Therefore, suchplaques are disseminated in both time and space.

It is believed that the pathogenesis involves a local disruption of theblood brain barrier which causes a localized immune and inflammatoryresponse, with consequent damage to myelin and hence to neurons.

Clinically, MS exists in both sexes and can occur at any age. However,its most common presentation is in the relatively young adult, oftenwith a single focal lesion such as a damage of the optic nerve, an areaof anesthesia (loss of sensation), or paraesthesia (localize loss offeeling), or muscular weakness. In addition, vertigo, double vision,localized pain, incontinence, and pain in the arms and legs may occurupon flexing of the neck, as well as a large variety of less commonsymptoms.

An initial attack of MS is often transient, and it may be weeks, months,or years before a further attack occurs. Some individuals may enjoy astable, relatively event free condition for a great number of years,while other less fortunate ones may experience a continual downhillcourse ending in complete paralysis. There is, most commonly, a seriesof remission and relapses, in which each relapse leaves a patientsomewhat worse than before. Relapses may be triggered by stressfulevents, viral infections or toxins. Therein, elevated body temperature,i.e., a fever, will make the condition worse, or as a reduction oftemperature by, for example, a cold bath, may make the condition better.

In yet another embodiment, a sirtuin-modulating compound that increasesthe level and/or activity of a sirtuin protein may be used to treattrauma to the nerves, including, trauma due to disease, injury(including surgical intervention), or environmental trauma (e.g.,neurotoxins, alcoholism, etc.).

Sirtuin-modulating compounds that increase the level and/or activity ofa sirtuin protein may also be useful to prevent, treat, and alleviatesymptoms of various PNS disorders, such as the ones described below. ThePNS is composed of the nerves that lead to or branch off from the spinalcord and CNS. The peripheral nerves handle a diverse array of functionsin the body, including sensory, motor, and autonomic functions. When anindividual has a peripheral neuropathy, nerves of the PNS have beendamaged. Nerve damage can arise from a number of causes, such asdisease, physical injury, poisoning, or malnutrition. These agents mayaffect either afferent or efferent nerves. Depending on the cause ofdamage, the nerve cell axon, its protective myelin sheath, or both maybe injured or destroyed.

The term “peripheral neuropathy” encompasses a wide range of disordersin which the nerves outside of the brain and spinal cord—peripheralnerves—have been damaged. Peripheral neuropathy may also be referred toas peripheral neuritis, or if many nerves are involved, the termspolyneuropathy or polyneuritis may be used.

Peripheral neuropathy is a widespread disorder, and there are manyunderlying causes. Some of these causes are common, such as diabetes,and others are extremely rare, such as acrylamide poisoning and certaininherited disorders. The most common worldwide cause of peripheralneuropathy is leprosy. Leprosy is caused by the bacterium Mycobacteriumleprae, which attacks the peripheral nerves of affected people.

Leprosy is extremely rare in the United States, where diabetes is themost commonly known cause of peripheral neuropathy. It has beenestimated that more than 17 million people in the United States andEurope have diabetes-related polyneuropathy. Many neuropathies areidiopathic; no known cause can be found. The most common of theinherited peripheral neuropathies in the United States isCharcot-Marie-Tooth disease, which affects approximately 125,000persons.

Another of the better known peripheral neuropathies is Guillain-Barrésyndrome, which arises from complications associated with viralillnesses, such as cytomegalovirus, Epstein-Barr virus, and humanimmunodeficiency virus (HIV), or bacterial infection, includingCampylobacter jejuni and Lyme disease. The worldwide incidence rate isapproximately 1.7 cases per 100,000 people annually. Other well-knowncauses of peripheral neuropathies include chronic alcoholism, infectionof the varicella-zoster virus, botulism, and poliomyelitis. Peripheralneuropathy may develop as a primary symptom, or it may be due to anotherdisease. For example, peripheral neuropathy is only one symptom ofdiseases such as amyloid neuropathy, certain cancers, or inheritedneurologic disorders. Such diseases may affect the PNS and the CNS, aswell as other body tissues.

Other PNS diseases treatable with sirtuin-modulating compounds thatincrease the level and/or activity of a sirtuin protein include:Brachial Plexus Neuropathies (diseases of the cervical and firstthoracic roots, nerve trunks, cords, and peripheral nerve components ofthe brachial plexus. Clinical manifestations include regional pain,paresthesia; muscle weakness, and decreased sensation in the upperextremity. These disorders may be associated with trauma, includingbirth injuries; thoracic outlet syndrome; neoplasms, neuritis,radiotherapy; and other conditions. See Adams et al., Principles ofNeurology, 6th ed, pp 1351-2); Diabetic Neuropathies (peripheral,autonomic, and cranial nerve disorders that are associated with diabetesmellitus). These conditions usually result from diabetic microvascularinjury involving small blood vessels that supply nerves (vasa nervorum).Relatively common conditions which may be associated with diabeticneuropathy include third nerve palsy; mononeuropathy; mononeuritismultiplex; diabetic amyotrophy; a painful polyneuropathy; autonomicneuropathy; and thoracoabdominal neuropathy (see Adams et al.,Principles of Neurology, 6th ed, p 1325); mononeuropathies (disease ortrauma involving a single peripheral nerve in isolation, or out ofproportion to evidence of diffuse peripheral nerve dysfunction).Mononeuritis multiplex refers to a condition characterized by multipleisolated nerve injuries. Mononeuropathies may result from a wide varietyof causes, including ischemia; traumatic injury; compression; connectivetissue diseases; cumulative trauma disorders; and other conditions;Neuralgia (intense or aching pain that occurs along the course ordistribution of a peripheral or cranial nerve); Peripheral NervousSystem Neoplasms (neoplasms which arise from peripheral nerve tissue).This includes neurofibromas; Schwannomas; granular cell tumors; andmalignant peripheral nerve sheath tumors (see DeVita Jr et al., Cancer:Principles and Practice of Oncology, 5th ed, pp 1750-1); and NerveCompression Syndromes (mechanical compression of nerves or nerve rootsfrom internal or external causes). These may result in a conductionblock to nerve impulses, due to, for example, myelin sheath dysfunction,or axonal loss. The nerve and nerve sheath injuries may be caused byischemia; inflammation; or a direct mechanical effect; Neuritis (ageneral term indicating inflammation of a peripheral or cranial nerve).Clinical manifestation may include pain; paresthesias; paresis; orhyperesthesia; Polyneuropathies (diseases of multiple peripheralnerves). The various forms are categorized by the type of nerve affected(e.g., sensory, motor, or autonomic), by the distribution of nerveinjury (e.g., distal vs. proximal), by nerve component primarilyaffected (e.g., demyelinating vs. axonal), by etiology, or by pattern ofinheritance.

In another embodiment, a sirtuin activating compound may be used totreat or prevent chemotherapeutic induced neuropathy. The sirtuinmodulating compounds may be administered prior to administration of thechemotherapeutic agent, concurrently with administration of thechemotherapeutic drug, and/or after initiation of administration of thechemotherapeutic drug. If the sirtuin activating compound isadministered after the initiation of administration of thechemotherapeutic drug, it is desirable that the sirtuin activatingcompound be administered prior to, or at the first signs, ofchemotherapeutic induced neuropathy.

Chemotherapy drugs can damage any part of the nervous system.Encephalopathy and myelopathy are fortunately very rare. Damage toperipheral nerves is much more common and can be a side effect oftreatment experienced by people with cancers, such as lymphoma. Most ofthe neuropathy affects sensory rather than motor nerves. Thus, thecommon symptoms are tingling, numbness or a loss of balance. The longestnerves in the body seem to be most sensitive hence the fact that mostpatients will report numbness or pins and needles in their hands andfeet.

The chemotherapy drugs which are most commonly associated withneuropathy, are the Vinca alkaloids (anti-cancer drugs originallyderived from a member of the periwinkle—the Vinca plant genus) and aplatinum-containing drug called Cisplatin. The Vinca alkaloids includethe drugs vinblastine, vincristine and vindesine. Many combinationchemotherapy treatments for lymphoma for example CHOP and CVP containvincristine, which is the drug known to cause this problem mostfrequently. Indeed, it is the risk of neuropathy that limits the dose ofvincristine that can be administered.

Studies that have been performed have shown that most patients will losesome reflexes in their legs as a result of treatment with vincristineand many will experience some degree of tingling (paresthesia) in theirfingers and toes. The neuropathy does not usually manifest itself rightat the start of the treatment but generally comes on over a period of afew weeks. It is not essential to stop the drug at the first onset ofsymptoms, but if the neuropathy progresses this may be necessary. It isvery important that patients should report such symptoms to theirdoctors, as the nerve damage is largely reversible if the drug isdiscontinued. Most doctors will often reduce the dose of vincristine orswitch to another form of Vinca alkaloid such as vinblastine orvindesine if the symptoms are mild. Occasionally, the nerves supplyingthe bowel are affected causing abdominal pain and constipation.

In another embodiment, a sirtuin activating compound may be used totreat or prevent a polyglutamine disease. Huntington's Disease (HD) andSpinocerebellar ataxia type 1 (SCA1) are just two examples of a class ofgenetic diseases caused by dynamic mutations involving the expansion oftriplet sequence repeats. In reference to this common mechanism, thesedisorders are called trinucleotide repeat diseases. At least 14 suchdiseases are known to affect human beings. Nine of them, including SCA1and Huntington's disease, have CAG as the repeated sequence (see Table 2below). Since CAG codes for an amino acid called glutamine, these ninetrinucleotide repeat disorders are collectively known as polyglutaminediseases.

Although the genes involved in different polyglutamine diseases havelittle in common, the disorders they cause follow a strikingly similarcourse. Each disease is characterized by a progressive degeneration of adistinct group of nerve cells. The major symptoms of these diseases aresimilar, although not identical, and usually affect people in midlife.Given the similarities in symptoms, the polyglutamine diseases arehypothesized to progress via common cellular mechanisms. In recentyears, scientists have made great strides in unraveling what themechanisms are.

Above a certain threshold, the greater the number of glutamine repeatsin a protein, the earlier the onset of disease and the more severe thesymptoms. This suggests that abnormally long glutamine tracts rendertheir host protein toxic to nerve cells.

To test this hypothesis, scientists have generated geneticallyengineered mice expressing proteins with long polyglutamine tracts.Regardless of whether the mice express full-length proteins or onlythose portions of the proteins containing the polyglutamine tracts, theydevelop symptoms of polyglutamine diseases. This suggests that a longpolyglutamine tract by itself is damaging to cells and does not have tobe part of a functional protein to cause its damage.

For example, it is thought that the symptoms of SCA1 are not directlycaused by the loss of normal ataxin-1 function but rather by theinteraction between ataxin-1 and another protein called LANP. LANP isneeded for nerve cells to communicate with one another and thus fortheir survival. When the mutant ataxin-1 protein accumulates insidenerve cells, it “traps” the LANP protein, interfering with its normalfunction. After a while, the absence of LANP function appears to causenerve cells to malfunction.

TABLE 2 Summary of Polyglutamine Diseases. Normal Disease GeneChromosomal Pattern of repeat repeat Disease name location inheritanceProtein length length Spinobulbar AR Xq13-21 X-linked androgen 9-3638-62 muscular recessive receptor atrophy (AR) (Kennedy disease)Huntington's HD 4p16.3 autosomal huntingtin 6-35  36-121 diseasedominant Dentatorubral- DRPLA 12p13.31 autosomal atrophin-1 6-35 49-88pallidoluysian dominant atrophy (Haw River syndrome) SpinocerebellarSCA1 6p23 autosomal ataxin-1 6-44 39-82 ataxia type 1 dominantSpinocerebellar SCA2 12q24.1 autosomal ataxin-2 15-31  36-63 ataxia type2 dominant Spinocerebellar SCA3 14q32.1 autosomal ataxin-3 12-40  55-84ataxia type 3 dominant (Machado- Joseph disease) Spinocerebellar SCA619p13 autosomal α1_(A)- 4-18 21-33 ataxia type 6 dominant voltage-dependent calcium channel subunit Spinocerebellar SCA7 3p12-13 autosomalataxin-7 4-35  37-306 ataxia type 7 dominant Spinocerebellar SCA17 6q27autosomal TATA 25-42  45-63 ataxia type 17 dominant binding protein

Many transcription factors have also been found in neuronal inclusionsin different diseases. It is possible that these transcription factorsinteract with the polyglutamine-containing proteins and then becometrapped in the neuronal inclusions. This in turn might keep thetranscription factors from turning genes on and off as needed by thecell. Another observation is hypoacetylation of histones in affectedcells. This has led to the hypothesis that Class I/II HistoneDeacetylase (HDAC I/II) inhibitors, which are known to increase histoneacetylation, may be a novel therapy for polyglutamine diseases (U.S.patent application Ser. No. 10/476,627; “Method of treatingneurodegenerative, psychiatric, and other disorders with deacetylaseinhibitors”).

In yet another embodiment, the invention provides a method for treatingor preventing neuropathy related to ischemic injuries or diseases, suchas, for example, coronary heart disease (including congestive heartfailure and myocardial infarctions), stroke, emphysema, hemorrhagicshock, peripheral vascular disease (upper and lower extremities) andtransplant related injuries.

In certain embodiments, the invention provides a method to treat acentral nervous system cell to prevent damage in response to a decreasein blood flow to the cell. Typically the severity of damage that may beprevented will depend in large part on the degree of reduction in bloodflow to the cell and the duration of the reduction. By way of example,the normal amount of perfusion to brain gray matter in humans is about60 to 70 mL/100 g of brain tissue/min. Death of central nervous systemcells typically occurs when the flow of blood falls below approximately8-10 mL/100 g of brain tissue/min, while at slightly higher levels (i.e.20-35 mL/100 g of brain tissue/min) the tissue remains alive but notable to function. In one embodiment, apoptotic or necrotic cell deathmay be prevented. In still a further embodiment, ischemic-mediateddamage, such as cytoxic edema or central nervous system tissue anoxemia,may be prevented. In each embodiment, the central nervous system cellmay be a spinal cell or a brain cell.

Another aspect encompasses administrating a sirtuin activating compoundto a subject to treat a central nervous system ischemic condition. Anumber of central nervous system ischemic conditions may be treated bythe sirtuin activating compounds described herein. In one embodiment,the ischemic condition is a stroke that results in any type of ischemiccentral nervous system damage, such as apoptotic or necrotic cell death,cytoxic edema or central nervous system tissue anoxia. The stroke mayimpact any area of the brain or be caused by any etiology commonly knownto result in the occurrence of a stroke. In one alternative of thisembodiment, the stroke is a brain stem stroke. Generally speaking, brainstem strokes strike the brain stem, which control involuntarylife-support functions such as breathing, blood pressure, and heartbeat.In another alternative of this embodiment, the stroke is a cerebellarstroke. Typically, cerebellar strokes impact the cerebellum area of thebrain, which controls balance and coordination. In still anotherembodiment, the stroke is an embolic stroke. In general terms, embolicstrokes may impact any region of the brain and typically result from theblockage of an artery by a vaso-occlusion. In yet another alternative,the stroke may be a hemorrhagic stroke. Like ischemic strokes,hemorrhagic stroke may impact any region of the brain, and typicallyresult from a ruptured blood vessel characterized by a hemorrhage(bleeding) within or surrounding the brain. In a further embodiment, thestroke is a thrombotic stroke. Typically, thrombotic strokes result fromthe blockage of a blood vessel by accumulated deposits.

In another embodiment, the ischemic condition may result from a disorderthat occurs in a part of the subject's body outside of the centralnervous system, but yet still causes a reduction in blood flow to thecentral nervous system. These disorders may include, but are not limitedto a peripheral vascular disorder, a venous thrombosis, a pulmonaryembolus, arrhythmia (e.g. atrial fibrillation), a myocardial infarction,a transient ischemic attack, unstable angina, or sickle cell anemia.Moreover, the central nervous system ischemic condition may occur asresult of the subject undergoing a surgical procedure. By way ofexample, the subject may be undergoing heart surgery, lung surgery,spinal surgery, brain surgery, vascular surgery, abdominal surgery, ororgan transplantation surgery. The organ transplantation surgery mayinclude heart, lung, pancreas, kidney or liver transplantation surgery.Moreover, the central nervous system ischemic condition may occur as aresult of a trauma or injury to a part of the subject's body outside thecentral nervous system. By way of example, the trauma or injury maycause a degree of bleeding that significantly reduces the total volumeof blood in the subject's body. Because of this reduced total volume,the amount of blood flow to the central nervous system is concomitantlyreduced. By way of further example, the trauma or injury may also resultin the formation of a vaso-occlusion that restricts blood flow to thecentral nervous system.

Of course it is contemplated that the sirtuin activating compounds maybe employed to treat the central nervous system ischemic conditionirrespective of the cause of the condition. In one embodiment, theischemic condition results from a vaso-occlusion. The vaso-occlusion maybe any type of occlusion, but is typically a cerebral thrombosis or anembolism. In a further embodiment, the ischemic condition may resultfrom a hemorrhage. The hemorrhage may be any type of hemorrhage, but isgenerally a cerebral hemorrhage or a subararachnoid hemorrhage. In stillanother embodiment, the ischemic condition may result from the narrowingof a vessel. Generally speaking, the vessel may narrow as a result of avasoconstriction such as occurs during vasospasms, or due toarteriosclerosis. In yet another embodiment, the ischemic conditionresults from an injury to the brain or spinal cord.

In yet another aspect, a sirtuin activating compound may be administeredto reduce infarct size of the ischemic core following a central nervoussystem ischemic condition. Moreover, a sirtuin activating compound mayalso be beneficially administered to reduce the size of the ischemicpenumbra or transitional zone following a central nervous systemischemic condition.

In one embodiment, a combination drug regimen may include drugs orcompounds for the treatment or prevention of neurodegenerative disordersor secondary conditions associated with these conditions. Thus, acombination drug regimen may include one or more sirtuin activators andone or more anti-neurodegeneration agents. For example, one or moresirtuin-activating compounds can be combined with an effective amount ofone or more of: L-DOPA; a dopamine agonist; an adenosine A₂A receptorantagonist; a COMT inhibitor; a MAO inhibitor; an N-NOS inhibitor; asodium channel antagonist; a selective N-methyl D-aspartate (NMDA)receptor antagonist; an AMPA/kainate receptor antagonist; a calciumchannel antagonist; a GABA-A receptor agonist; an acetyl-cholineesterase inhibitor; a matrix metalloprotease inhibitor; a PARPinhibitor; an inhibitor of p38 MAP kinase or c-jun-N-terminal kinases;TPA; NDA antagonists; beta-interferons; growth factors; glutamateinhibitors; and/or as part of a cell therapy.

Exemplary N-NOS inhibitors include4-(6-amino-pyridin-2-yl)-3-methoxyphenol6-[4-(2-dimethylamino-ethoxy)-2-methoxy-phenyl]-pyridin-2-yl-amine,6-[4-(2-dimethylamino-ethoxy)-2,3-dimet-hyl-phenyl]-pyridin-2-yl-amine,6-[4-(2-pyrrolidinyl-ethoxy)-2,3-dimethyl-p-henyl]-pyridin-2-yl-amine,6-[4-(4-(n-methyl)piperidinyloxy)-2,3-dimethyl-p-henyl]-pyridin-2-yl-amine,6-[4-(2-dimethylamino-ethoxy)-3-methoxy-phenyl]-pyridin-2-yl-amine,6-[4-(2-pyrrolidinyl-ethoxy)-3-methoxy-phenyl]-pyridin-2-yl-amine,6-{4-[2-(6,7-dimethoxy-3,4-dihydro-1h-isoquinolin-2-yl)-ethoxy]-3-methoxy-phenyl}-pyridin-2-yl-amine,6-{3-methoxy-4-[2-(4-phenethyl-piper-azin-1-yl)-ethoxy]-phenyl}-pyridin-2-yl-amine,6-{3-methoxy-4-[2-(4-methyl-piperazin-1-yl)-ethoxy]-phenyl}-pyridin-2-yl-amine,6-{4-[2-(4-dimethylamin-o-piperidin-1-yl)-ethoxy]-3-methoxy-phenyl}-pyridin-2-yl-amine,6-[4-(2-dimethylamino-ethoxy)-3-ethoxy-phenyl]-pyridin-2-yl-amine,6-[4-(2-pyrrolidinyl-ethoxy)-3-ethoxy-phenyl]-pyridin-2-yl-amine,6-[4-(2-dimethylamino-ethoxy)-2-isopropyl-phenyl]-pyridin-2-yl-amine,4-(6-amino-pyridin-yl)-3-cyclopropyl-phenol6-[2-cyclopropyl-4-(2-dimethy-lamino-ethoxy)-phenyl]-pyridin-2-yl-amine,6-[2-cyclopropyl-4-(2-pyrrolidin-1-yl-ethoxy)-phenyl]-pyridin-2-yl-amine,3-[3-(6-amino-pyridin-2-yl)-4-cycl-opropyl-phenoxy]-pyrrolidine-1-carboxylicacid tert-butyl ester6-[2-cyclopropyl-4-(1-methyl-pyrrolidin-3-yl-oxy)-phenyl]-pyridin-2-yl-amine,4-(6-amino-pyridin-2-yl)-3-cyclobutyl-phenol6-[2-cyclobutyl-4-(2-dime-thylamino-ethoxy)-phenyl]-pyridin-2-yl-amine,6-[2-cyclobutyl-4-(2-pyrrolid-in-1-yl-ethoxy)-phenyl]-pyridin-2-yl-amine,6-[2-cyclobutyl-4-(1-methyl-pyr-rolidin-3-yl-oxy)-phenyl]-pyridin-2-yl-amine,4-(6-amino-pyridin-2-yl)-3-cy-clopentyl-phenol6-[2-cyclopentyl-4-(2-dimethylamino-ethoxy)-phenyl]-pyrid-in-2-yl-amine,6-[2-cyclopentyl-4-(2-pyrrolidin-1yl-ethoxy)-phenyl]-pyridin-2-yl-amine,3-[4-(6-amino-pyridin-2yl)-3-methoxy-phenoxy]-pyrrolidine-1-ca-rboxylicacid tert butyl ester6-[4-(1-methyl-pyrrolidin-3-yl-oxy)-2-metho-xy-phenyl]-pyridin-2-yl-amine,4-[4-(6-amino-pyridin-2-yl)-3-methoxy-phenoxy-]-piperidine-1-carboxylicacid tert butyl ester6-[2-methoxy-4-(1-methyl-piperidin-4-yl-oxy)-phenyl]-pyridin-2-yl-amine,6-[4-(allyloxy)-2-methoxy-ph-enyl]-pyridin-2-yl-amine,4-(6-amino-pyridin-2-yl)-3-methoxy-6-allyl-phenol 12 and4-(6-amino-pyridin-2-yl)-3-methoxy-2-allyl-phenol 134-(6-amino-pyridin-2-yl)-3-methoxy-6-propyl-phenol6-[4-(2-dimethylamino-ethoxy)-2-methoxy-5-propyl-phenyl]-pyridin-yl-amine,6-[2-isopropyl-4-(pyrrolidin-3-yl-oxy)-phenyl]-pyridin-2-yl-amine,6-[2-isopropyl-4-(piperidin-3-yl-oxy)-phenyl]-pyridin-2-yl-amine,6-[2-isopropyl-4-(1-methyl-azetidin-3-yl-oxy)-phenyl]-pyridin-2-yl-amine,6-[2-isopropyl-4-(1-methyl-piperidin-4-yl-oxy)-phenyl]-pyridin-2-yl-amine,6-[2-isopropyl-4-(1-methyl-pyrrolidin-3-yl-oxy)-phenyl]-pyridin-2-yl-amin-e6-[2-isopropyl-4-(1-methyl-pyrrolidin-3-yl-oxy)-phenyl]-pyridin-2-yl-amine,6-[2-isopropyl-4-(2-methyl-2-aza-bicyclo[2.2.1]hept-5-yl-oxy)-phenyl]-p-yridin-2-yl-amine,6-[4-(2-dimethylamino-ethoxy)-2-methoxy-phenyl]-pyridin-2-yl-amine,6-{4-[2-(benzyl-methyl-amino)-ethoxy]-2-methoxy-phenyl}-pyridin-2-yl-amine,6-[2-methoxy-4-(2-pyrrolidin-1-yl-ethoxy)-phenyl]-pyridin-2-yl-amine,2-(6-amino-pyridin-2-yl)-5-(2-dimethylamino-ethoxy)-phenol2-[4-(6-amino-pyridin-2-yl)-3-methoxy-phenoxy]-acetamide6-[4-(2-amino-ethoxy)-2-methoxy-phenyl]-pyridin-2-yl-amine,6-{4-[2-(3,4-dihydro-1h-isoquinolin-2-yl)-ethoxy]-2-methoxy-phenyl}-pyrid-in-2-yl-amine,2-[4-(6-amino-pyridin-2-yl)-3-methoxy-phenoxy]-ethanol6-{2-methoxy-4-[2-(2,2,6,6-tetramethyl-piperidin-1-yl)-ethoxy]-phenyl}-py-ridin-2-yl-amine,6-{4-[2-(2,5-dimethyl-pyrrolidin-1-yl)-ethoxy]-2-methoxy-phenyl}-pyridin-2-yl-amine,6-{4-[2-(2,5-dimethyl-pyrrolidin-1-yl)-ethoxy]-2-methoxy-phenyl}-pyridin-2-yl-amine,2-[4-(6-amino-pyridin-2-yl)-3-methoxy-phenoxy]-1-(2,2,6,6-tetramethyl-piperidin-1-yl)-ethanone6-[2-methoxy-4-(1-methyl-pyrrolidin-2-yl-methoxy)-phenyl]-pyridin-2-yl-amine,6-[4-(2-dimethylamino-ethoxy)-2-propoxy-phenyl]-pyridin-2-yl-amine,6-{4-[2-(benzyl-methyl-amino)-ethoxy]-2-propoxy-phenyl}-pyridin-2-yl-amin-e6-[4-(2-ethoxy-ethoxy)-2-methoxy-phenyl]-pyridin-2-yl-amine,6-[4-(2-dimethylamino-ethoxy)-2-isopropoxy-phenyl]-pyridin-2-yl-amine,6-[4-(2-ethoxy-ethoxy)-2-isopropoxy-phenyl]-pyridin-2-yl-amine,6-[2-methoxy-4-(3-methyl-butoxy)-phenyl]-pyridin-2-yl-amine,6-[4-(2-dimethylamino-ethoxy)-2-ethoxy-phenyl]-pyridin-2-yl-amine,6-{4-[2-(benzyl-methyl-amino)-ethoxy]-2-ethoxy-phenyl}-pyridin-2-yl-amine,6-[2-ethoxy-4-(3-methyl-butoxy)-phenyl]-pyridin-2-yl-amine,1-(6-amino-3-aza-bicyclo[3.1.0]hex-3-yl)-2-[4-(6-amino-pyridin-2-yl)-3-et-hoxy-phenoxy]-ethanone6-[2-ethoxy-4-(2-pyrrolidin-1-yl-ethoxy)-phenyl]-py-ridin-2-yl-amine,3-{2-[4-(6-amino-pyridin-2-yl)-3-ethoxy-phenoxy]-ethyl}-3-aza-bicyclo[3.1.0]hex-6-yl-amine,1-(6-amino-3-aza-bicyclo[3.1.0]hex-3-yl)-2-[4-(6-amino-pyridin-2-yl)-3-methoxy-phenoxy]-ethanone3-{2-[4-(6-amino-pyridin-2-yl)-3-methoxy-phenoxy]-ethyl}-3-aza-bicyclo[3.1.0]hex-6-yl-amine,6-[2-isopropoxy-4-(2-pyrrolidin-1-yl-ethoxy)-phenyl]-py-ridin-2-yl-amine,6-{4-[2-(benzyl-methyl-amino)-ethoxy]-2-isopropoxy-phenyl-}-pyridin-2-yl-amine,6-[4-(2-dimethylamino-ethoxy)-2-methoxy-5-propyl-phen-yl]-pyridin-2-yl-amine,6-[5-allyl-4-(2-dimethylamino-ethoxy)-2-methoxy-phe-nyl]-pyridin-2-yl-amine,6-[5-allyl-2-methoxy-4-(2-pyrrolidin-1-yl-ethoxy)-phenyl]-pyridin-2-yl-amine,6-[3-allyl-4-(2-dimethylamino-ethoxy)-2-methoxy-phenyl]-pyridin-2-yl-amine,6-[2-methoxy-4-(pyrrolidin-3-yl-oxy)-phenyl]-p-yridin-2-yl-amine,6-[2-methoxy-4-(1-methyl-pyrrolidin-3-yl-oxy)-phenyl]-py-ridin-2-yl-amine,6-[2-ethoxy-4-(pyrrolidin-3-yl-oxy)-phenyl]-pyridin-2-yl-amine,6-[2-isopropoxy-4-(pyrrolidin-3-yl-oxy)-phenyl]-pyridin-2-yl-amine,6-[2-methoxy-4-(piperidin-4-yl-oxy)-phenyl]-pyridin-2-yl-amine,6-[2-methoxy-4-(2,2,6,6-tetramethyl-piperidin-4-yl-oxy)-phenyl]-pyridin-2-yl-amine,6-[2-isopropoxy-4-(pyrrolidin-3-yl-oxy)-phenyl]-pyridin-2-yl-amine,3-[4-(6-amino-pyridin-2-yl)-3-methoxy-phenoxy]-azetidine-1-carboxylicacid tert-butyl ester6-[4-(azetidin-3-yl-oxy)-2-methoxy-phenyl]-pyridin-2-yl-amine,6-[2-methoxy-4-(1-methyl-azetidin-3-yl-oxy)-phenyl]-pyridin-2-y-l-amine,6-[2-isopropoxy-4-(pyrrolidin-3-yl-oxy)-phenyl]-pyridin-2-yl-amine,6-[2-isopropoxy-4-(pyrrolidin-3-yl-oxy)-phenyl]-pyridin-2-yl-amine,6-[2-methoxy-4-(pyrrolidin-3-yl-oxy)-phenyl]-pyridin-2-yl-amine,6-[2-methoxy-4-(1-methyl-pyrrolidin-3-yl-oxy)-phenyl]-pyridin-2-yl-amine,6-[2-methoxy-4-(1-methyl-pyrrolidin-3-yl-oxy)-phenyl]-pyridin-2-yl-amine,6-[2-methoxy-4-(2-methyl-2-aza-bicyclo[2.2.1]hept-5-yl-oxy)-phenyl]-pyrid-in-2-yl-amine,6-[2-methoxy-4-(1-methyl-piperidin-4-yl-oxy)-phenyl]-pyridin-2-yl-amine,6-[4-(1-ethyl-piperidin-4-yl-oxy)-2-methoxy-phenyl]-pyridin-2-yl-amine,6-[5-allyl-2-methoxy-4-(1-methyl-pyrrolidin-3-yl-oxy)-phenyl]-pyr-idin-2-yl-amine,6-[4-(2-dimethylamino-ethoxy)-2,6-dimethyl-phenyl]-pyridin-2-yl-amine,6-[2,6-dimethyl-4-(3-piperidin-1-yl-propoxy)-phenyl]-pyridin-2-yl-amine,6-[2,6-dimethyl-4-(2-pyrrolidin-1-yl-ethoxy)-phenyl]-pyridin-2-y-l-amine,6-{2,6-dimethyl-4-[3-(4-methyl-piperazin-1-yl)-propoxy]-phenyl}-py-ridin-2-yl-amine,6-[2,6-dimethyl-4-(2-morpholin-4-yl-ethoxy)-phenyl]-pyrid-in-2-yl-amine,6-{4-[2-(benzyl-methyl-amino)-ethoxy]-2,6-dimethyl-phenyl}-p-yridin-2-yl-amine,2-[4-(6-amino-pyridin-2-yl)-3,5-dimethyl-phenoxy]-acetam-ide6-[4-(2-amino-ethoxy)-2,6-dimethyl-phenyl]-pyridin-2-yl-amine,6-[2-isopropyl-4-(2-pyrrolidin-1-yl-ethoxy)-phenyl]-pyridin-2-yl-amine,2-(2,5-dimethyl-pyrrolidin-1-yl)-6-[2-isopropyl-4-(2-pyrrolidin-1-yl-etho-xy)-phenyl]-pyridine6-{4-[2-(3,5-dimethyl-piperidin-1-yl)-ethoxy]-2-isopr-opyl-phenyl}-pyridin-2-yl-amine,6-[4-(2-dimethylamino-ethoxy)-2-isopropyl-phenyl]-pyridin-2-yl-amine,6-[2-tert-butyl-4-(2-dimethylamino-ethoxy)-phen-yl]-pyridin-2-yl-amine,6-[2-tert-butyl-4-(2-pyrrolidin-1-yl-ethoxy)-phenyl-]-pyridin-2-yl-amine,6-[4-(2-pyrrolidinyl-ethoxy)-2,5-dimethyl-phenyl]-pyr-idin-2-yl-amine,6-[4-(2-dimethylamino-ethoxy)-2,5-dimethyl-phenyl]-pyridin-2-yl-amine,6-[4-(2-(4-phenethylpiperazin-1-yl)-ethoxy)-2,5-dimethyl-pheny-l]-pyridin-2-yl-amine,6-[2-cyclopropyl-4-(2-dimethylamino-1-methyl-ethoxy)-phenyl]-pyridin-2-yl-amine,6-[cyclobutyl-4-(2-dimethylamino-1-methyl-etho-xy)-phenyl]-pyridin-2-yl-amine,6-[4-(allyloxy)-2-cyclobutyl-phenyl]-pyridi-n-2-ylamine,2-allyl-4-(6-amino-pyridin-2-yl)-3-cyclobutyl-phenol and2-allyl-4-(6-amino-pyridin-2-yl)-5-cyclobutyl-phenol4-(6-amino-pyridin-2yl)-5-cyclobutyl-2-propyl-phenol4-(6-amino-pyridin-2-yl)-3-cyclobutyl-2-propyl-phenol6-[2-cyclobutyl-4-(2-dimethylamino-1-methyl-ethoxy)-5-propyl-phenyl]-pyri-din-2-yl-amine,6-[2-cyclobutyl-4-(2-dimethylamino-1-methyl-ethoxy)-3-propy-1-phenyl]-pyridin-2-yl-amine,6-[2-cyclobutyl-4-(2-dimethylamino-ethoxy)-5-propyl-phenyl]-pyridin-2-yl-amine,6-[2-cyclobutyl-4-(2-dimethylamino-ethox-y)-3-propyl-phenyl]-pyridin-2-yl-amine,6-[2-cyclobutyl-4-(1-methyl-pyrroli-din-3-yl-oxy)-5-propyl-phenyl]-pyridin-2-yl-amine,6-[cyclobutyl-4-(1-methyl-1-pyrrolidin-3-yl-oxy)-3-propyl-phenyl]-pyridin-2-yl-amine,2-(4-benzyloxy-5-hydroxy-2-methoxy-phenyl)-6-(2,5-dimethyl-pyrrol-1-yl)-p-yridine6-[4-(2-dimethylamino-ethoxy)-5-ethoxy-2-methoxy-phenyl]-pyridin-2-yl-amine,6-[5-ethyl-2-methoxy-4-(1-methyl-piperidin-4-yl-oxy)-phenyl]-pyr-idin-2-yl-amine,6-[5-ethyl-2-methoxy-4-(piperidin-4-yl-oxy)-phenyl]-pyridi-n-2-yl-amine,6-[2,5-dimethoxy-4-(1-methyl-pyrrolidin-3-yl-oxy)-phenyl]-pyr-idin-2-yl-amine,6-[4-(2-dimethylamino-ethoxy)-5-ethyl-2-methoxy-phenyl]-py-ridin-2-yl-amine.

Exemplary NMDA receptor antagonist include(+)-(1S,2S)-1-(4-hydroxy-phenyl)-2-(4-hydroxy-4-phenylpiperidino)-1-pro-panol,(1S,2S)-1-(4-hydroxy-3-methoxyphenyl)-2-(4-hydroxy-4-phenylpiperi-dino)-1-propanol,(3R,4S)-3-(4-(4-fluorophenyl)-4-hydroxypiperidin-1-yl-)-chroman-4,7-diol,(1R*,2R*)-1-(4-hydroxy-3-methylphenyl)-2-(4-(4-fluoro-phenyl)-4-hydroxypiperidin-1-yl)-propan-1-ol-mesylateor a pharmaceutically acceptable acid addition salt thereof.

Exemplary dopamine agonist include ropininole; L-dopa decarboxylaseinhibitors such as carbidopa or benserazide, bromocriptine,dihydroergocryptine, etisulergine, AF-14, alaptide, pergolide,piribedil; dopamine DI receptor agonists such as A-68939, A-77636,dihydrexine, and SKF-38393; dopamine D2 receptor agonists such ascarbergoline, lisuride, N-0434, naxagolide, PD-18440, pramipexole,quinpirole and ropinirole; dopamine/β-adrenegeric receptor agonists suchas DPDMS and dopexamine; dopamine/5-HT uptake inhibitor/5-HT-1A agonistssuch as roxindole; dopamine/opiate receptor agonists such as NIH-10494;α2-adrenergic antagonist/dopamine agonists such as terguride;α2-adrenergic antagonist/dopamine D2 agonists such as ergolines andtalipexole; dopamine uptake inhibitors such as GBR-12909, GBR-13069,GYKI-52895, and NS-2141; monoamine oxidase-B inhibitors such asselegiline, N-(2-butyl)-N-methylpropargylamine,N-methyl-N-(2-pentyl)propargylamine, AGN-1133, ergot derivatives,lazabemide, LU-53439, MD-280040 and mofegiline; and COMT inhibitors suchas CGP-28014.

Exemplary acetyl cholinesterase inhibitors include donepizil,1-(2-methyl-1H-benzimida-zol-5-yl)-3-[1-(phenylmethyl)-4-piperidinyl]-1-propanone;1-(2-phenyl-1H-benzimidazol-5-yl)-3-[1-(phenylmethyl)-4-piperidinyl]-1-pr-opanone;1-(1-ethyl-2-methyl-1H-benzimidazol-5-yl)-3-[1-(phenylmethyl)-4-piperidinyl]-1-propanone;1-(2-methyl-6-benzothiazolyl)-3-[1-(phenylmethyl)-4-piperidinyl]-1-propanone;1-(2-methyl-6-benzothiazolyl)-3-[1-[(2-methyl-4-thiazolyl)methyl]-4-piperidinyl]-1-propanone;1-(5-methyl-benzo[b]thie-n-2-yl)-3-[1-(phenylmethyl)-4-piperidinyl]-1-propanone;1-(6-methyl-benzo[b]thien-2-yl)-3-[1-(phenylmethyl)-4-piperidinyl]-1-prop-anone;1-(3,5-dimethyl-benzo[b]thien-2-yl)-3-[1-(phenylmethyl)-4-piperidin-yl]-1-propanone;1-(benzo[b]thien-2-yl)-3-[1-(phenylmethyl)-4-piperidinyl]-1-propanone;1-(benzofuran-2-yl)-3-[1-(phenylmethyl)-4-piperidinyl]-1-pro-panone;1-(1-phenylsulfonyl-6-methyl-indol-2-yl)-3-[1-(phenylmethyl)-4-pip-eridinyl]-1-propanone;1-(6-methyl-indol-2-yl)-3-[1-(phenylmethyl)-4-piper-idinyl]-1-propanone;1-(1-phenylsulfonyl-5-amino-indol-2-yl)-3-[1-(phenylm-ethyl)-4-piperidinyl]-1-propanone;1-(5-amino-indol-2-yl)-3-[1-(phenylmet-hyl)-4-piperidinyl]-1-propanone;and1-(5-acetylamino-indol-2-yl)-3-[1-(ph-enylmethyl)-4-piperidinyl]-1-propanone.1-(6-quinolyl)-3-[1-(phenylmethyl)-4-piperidinyl]-1-propanone;1-(5-indolyl)-3-[1-(phenylmethyl)-4-piperidiny-l]-1-propanone;1-(5-benzthienyl)-3-[1-(phenylmethyl)-4-piperidinyl]-1-pro-panone;1-(6-quinazolyl)-3-[1-(phenylmethyl)-4-piperidinyl]-1-propanone;1-(6-benzoxazolyl)-3-[1-(phenylmethyl)-4-piperidinyl]-1-propanone;1-(5-benzofuryl)-3-[1-(phenylmethyl)-4-piperidinyl]-1-propanone;1-(5-methyl-benzimidazol-2-yl)-3-[1-(phenylmethyl)-4-piperidinyl]-1-propa-none;1-(6-methyl-benzimidazol-2-yl)-3-[1-(phenylmethyl)-4-piperidinyl]-1-propanone;1-(5-chloro-benzo[b]thien-2-yl)-3-[1-(phenylmethyl)-4-piperidin-yl]-1-propanone;1-(5-azaindol-2-yl)-3-[1-(phenylmethyl)-4-piperidinyl]-1-p-ropanone;1-(6-azabenzo[b]thien-2-yl)-3-[1-(phenylmethyl)-4-piperidinyl]-1-propanone;1-(1H-2-oxo-pyrrolo[2′,3′,5,6]benzo[b]thieno-2-yl)-3-[1-(phenylmethyl)-4-piperidinyl]-1-propanone;1-(6-methyl-benzothiazol-2-yl)-3-[1-(phenylmethyl)-4-piperidinyl]-1-propanone;1-(6-methoxy-indol-2-yl)-3-[1-(phenylmethyl)-4-piperidinyl]-1-propanone;1-(6-methoxy-benzo[b]thien-2-yl)-3-[1-(phenylmethyl)-4-piperidinyl]-1-pro-panone;1-(6-acetylamino-benzo[b]thien-2-yl)-3-[1-(phenylmethyl)-4-piperid-inyl]-1-propanone;1-(5-acetylamino-benzo[b]thien-2-yl)-3-[1-(phenylmethyl-)-4-piperidinyl]-1-propanone;6-hydroxy-3-[2-[1-(phenylmethyl)-4-piperidin-yl]ethyl]-1,2-benzisoxazole;5-methyl-3-[2-[1-(phenylmethyl)-4-piperidinyl-]ethyl]-1,2-benzisoxazole;6-methoxy-3 [2-[1(phenylmethyl)-4-piperidinyl]et-hyl]-1,2-benzisoxazole;6-acetamide-3-[2-[1-(phenylmethyl)-4-piperidinyl]-ethyl]-1,2-benzisoxazole;6-amino-3-[2-[1-(phenymethyl)-4-piperidinyl]ethy-l]-1,2-benzisoxazole;6-(4-morpholinyl)-3-[2-[1-(phenylmethyl)-4-piperidin-yl]ethyl]-1,2-benzisoxazole;5,7-dihydro-3-[2-[1-(phenylmethyl)-4-piperidi-nyl]ethyl]-6H-pyrrolo[4,5-f]-1,2-benzisoxazol-6-one;3-[2-[1-(phenylmethyl)-4-piperidinyl]ethyl]-1,2-benzisothiazole;3-[2-[1-(phenylmethyl)-4-piperidinyl]ethenyl]-1,2-benzisoxazole;6-phenylamino-3-[2-[1-(phenylmethyl)-4-piperidinyl]ethyl]-1,2,-benzisoxaz-ole;6-(2-thiazoly)-3-[2-[1-(phenylmethyl)-4-piperidinyl]ethyl]-1,2-benzis-oxazole;6-(2-oxazolyl)-3-[2-[1-(phenylmethyl)-4-piperidinyl]ethyl]-1,2-be-nzisoxazole;6-pyrrolidinyl-3-[2-[1-(phenylmethyl)-4-piperidinyl]ethyl]-1,2-benzisoxazole;5,7-dihydro-5,5-dimethyl-3-[2-[1-(phenylmethyl)-4-piperid-inyl]ethyl]-6H-pyrrolo[4,5-f]-1,2-benzisoxazole-6-one;6,8-dihydro-3-[2-[1-(phenylmethyl)-4-piperidinyl]ethyl]-7H-pyrrolo[5,4-g]-1,2-benzisoxazole-7-one;3-[2-[1-(phenylmethyl)-4-piperidinyl]ethyl]-5,6,-8-trihydro-7H-isoxazolo[4,5-g]-quinolin-7-one;1-benzyl-4-((5,6-dimethoxy-1-indanon)-2-yl)methylpiperidine,1-benzyl-4-((5,6-dimethoxy-1-indanon)-2-ylidenyl)methylpiperidine,1-benzyl-4-((5-methoxy-1-indanon)-2-yl)methylppiperidine,1-benzyl-4-((5,6-diethoxy-1-indanon)-2-yl)methylpiperidine,1-benzyl-4-((5,6-methnylenedioxy-1-indanon)-2-yl)methylpiperidine,1-(m-nitrobenzyl)-4-((5,6-dimethoxy-1-indanon)-2-yl)methylpiperidine,1-cyclohexymethyl-4-((5,6-dimethoxy-1-indanon)-2-yl)methylpiperidine,1-(m-florobenzyl)-4-((5,6-dimethoxy-1-indanon)-2-yl)methylpiperidine,1-benzyl-4-((5,6-dimethoxy-1-indanon)-2-yl)propylpiperidine, and1-benzyl-4-((5-isopropoxy-6-methoxy-1-indanon)-2-yl)methylpiperidine.

Exemplary calcium channel antagonists include diltiazem, omega-conotoxinGVIA, methoxyverapamil, amlodipine, felodipine, lacidipine, andmibefradil.

Exemplary GABA-A receptor modulators include clomethiazole; IDDB;gaboxadol (4,5,6,7-tetrahydroisoxazolo[5,4-c]pyridin-3-ol); ganaxolone(3α-hydroxy-3β-methyl-5α-pregnan-20-one); fengabine(2-[(butylimino)-(2-chlorophenyl)methyl]-4-chlorophenol);2-(4-methoxyphenyl)-2,5,6,7,8,9-hexahydro-pyrazolo[4,3-c]cinnolin-3-one;7-cyclobutyl-6-(2-methyl-2H-1,2,4-triazol-3-ylmethoxy)-3-phenyl-1,2,4-triazolo[4,3-b]pyridazine;(3-fluoro-4-methylphenyl)-N-({-1-[(2-methylphenyl)methyl]-benzimidazol-2-yl}methyl)-N-pentylcarboxamide;and 3-(aminomethyl)-5-methylhexanoic acid.

Exemplary potassium channel openers include diazoxide, flupirtine,pinacidil, levcromakalim, rilmakalim, chromakalim, PCO-400 and SKP450(2-[2″(1″,3″-dioxolone)-2-methyl]-4-(2′-oxo-1′-pyrrolidinyl)-6-nitro-2H-1-benzopyra-n).

Exemplary AMPA/kainate receptor antagonists include6-cyano-7-nitroquinoxalin-2,3-di-one (CNQX);6-nitro-7-sulphamoylbenzo[f]quinoxaline-2,3-dione (NBQX);6,7-dinitroquinoxaline-2,3-dione (DNQX);1-(4-aminophenyl)-4-methyl-7,8-m-ethylenedioxy-5H-2,3-benzodiazepinehydrochloride; and2,3-dihydroxy-6-nitro-7-sulfamoylbenzo-[f]quinoxaline.

Exemplary sodium channel antagonists include ajmaline, procainamide,flecamide and riluzole.

Exemplary matrix-metalloprotease inhibitors include4-[4-(4-fluorophenoxy)benzenesulfon-ylamino]tetrahydropyran-4-carboxylicacid hydroxyamide;5-Methyl-5-(4-(4′-fluorophenoxy)-phenoxy)-pyrimidine-2,4,6-trione;5-n-Butyl-5-(4-(4′-fluorophenoxy)-phenoxy)-pyrimidine-2,4,6-trione andprinomistat.

Poly(ADP ribose) polymerase (PARP) is an abundant nuclear enzyme whichis activated by DNA strand single breaks to synthesize poly (ADP ribose)from NAD. Under normal conditions, PARP is involved in base excisionrepair caused by oxidative stress via the activation and recruitment ofDNA repair enzymes in the nucleus. Thus, PARP plays a role in cellnecrosis and DNA repair. PARP also participates in regulating cytokineexpression that mediates inflammation. Under conditions where DNA damageis excessive (such as by acute excessive exposure to a pathologicalinsult), PARP is over-activated, resulting in cell-based energeticfailure characterized by NAD depletion and leading to ATP consumption,cellular necrosis, tissue injury, and organ damage/failure. PARP isthought to contribute to neurodegeneration by depleting nicotinamideadenine dinucleotide (NAD+) which then reduces adenosine triphosphate(ATP; Cosi and Marien, Ann. N.Y. Acad. Sci., 890:227, 1999) contributingto cell death which can be prevented by PARP inhibitors. Exemplory PARPinhibitors can be found in Southan and Szabo, Current MedicinalChemistry, 10:321, 2003.

Exemplary inhibitors of p38 MAP kinase and c-jun-N-terminal kinasesinclude pyridyl imidazoles, such as PD 169316, isomeric PD 169316, SB203580, SB 202190, SB 220026, and RWJ 67657. Others are described inU.S. Pat. No. 6,288,089, and incorporated by reference herein.

In an exemplary embodiment, a combination therapy for treating orpreventing MS comprises a therapeutically effective amount of one ormore sirtuin-modulating compounds that increase the level and/oractivity of a sirtuin protein and one or more of Avonex® (interferonbeta-1a), Tysabri® (natalizumab), or Fumaderm® (BG-12/Oral Fumarate).

In another embodiment, a combination therapy for treating or preventingdiabetic neuropathy or conditions associated therewith comprises atherapeutically effective amount of one or more sirtuin-modulatingcompounds that increase the level and/or activity of a sirtuin proteinand one or more of tricyclic antidepressants (TCAs) (including, forexample, imipramine, amytriptyline, desipramine and nortriptyline),serotonin reuptake inhibitors (SSRIs) (including, for example,fluoxetine, paroxetine, sertralene, and citalopram) and antiepilepticdrugs (AEDS) (including, for example, gabapentin, carbamazepine, andtopimirate).

In another embodiment, the invention provides a method for treating orpreventing a polyglutamine disease using a combination comprising atleast one sirtuin activating compound and at least one HDAC I/IIinhibitor. Examples of HDAC I/II inhibitors include hydroxamic acids,cyclic peptides, benzamides, short-chain fatty acids, and depudecin.

Examples of hydroxamic acids and hydroxamic acid derivatives, but arenot limited to, trichostatin A (TSA), suberoylanilide hydroxamic acid(SAHA), oxamflatin, suberic bishydroxamic acid (SBHA),m-carboxy-cinnamic acid bishydroxamic acid (CBHA), valproic acid andpyroxamide. TSA was isolated as an antifungi antibiotic (Tsuji et al(1976) J. Antibiot (Tokyo) 29:1-6) and found to be a potent inhibitor ofmammalian HDAC (Yoshida et al. (1990) J. Biol. Chem. 265:17174-17179).The finding that TSA-resistant cell lines have an altered HDAC evidencesthat this enzyme is an important target for TSA. Other hydroxamicacid-based HDAC inhibitors, SAHA, SBHA, and CBHA are synthetic compoundsthat are able to inhibit HDAC at micromolar concentration or lower invitro or in vivo. Glick et al. (1999) Cancer Res. 59:4392-4399. Thesehydroxamic acid-based HDAC inhibitors all possess an essentialstructural feature: a polar hydroxamic terminal linked through ahydrophobic methylene spacer (e.g. 6 carbon at length) to another polarsite which is attached to a terminal hydrophobic moiety (e.g., benzenering). Compounds developed having such essential features also fallwithin the scope of the hydroxamic acids that may be used as HDACinhibitors.

Cyclic peptides used as HDAC inhibitors are mainly cyclic tetrapeptides.Examples of cyclic peptides include, but are not limited to, trapoxin A,apicidin and depsipeptide. Trapoxin A is a cyclic tetrapeptide thatcontains a 2-amino-8-oxo-9,10-epoxy-decanoyl (AOE) moiety. Kijima et al.(1993) J. Biol. Chem. 268:22429-22435. Apicidin is a fungal metabolitethat exhibits potent, broad-spectrum antiprotozoal activitity andinhibits HDAC activity at nanomolar concentrations. Darkin-Rattray etal. (1996) Proc. Natl. Acad. Sci. USA. 93; 13143-13147. Depsipeptide isisolated from Chromobacterium violaceum, and has been shown to inhibitHDAC activity at micromolar concentrations.

Examples of benzamides include but are not limited to MS-27-275. Saitoet al. (1990) Proc. Natl. Acad. Sci. USA. 96:4592-4597. Examples ofshort-chain fatty acids include but are not limited to butyrates (e.g.,butyric acid, arginine butyrate and phenylbutyrate (PB)). Newmark et al.(1994) Cancer Lett. 78:1-5; and Carducci et al. (1997) Anticancer Res.17:3972-3973. In addition, depudecin which has been shown to inhibitHDAC at micromolar concentrations (Kwon et al. (1998) Proc. Natl. Acad.Sci. USA. 95:3356-3361) also falls within the scope of histonedeacetylase inhibitor as described herein.

Blood Coagulation Disorders

In other aspects, sirtuin-modulating compounds that increase the leveland/or activity of a sirtuin protein can be used to treat or preventblood coagulation disorders (or hemostatic disorders). As usedinterchangeably herein, the terms “hemostasis”, “blood coagulation,” and“blood clotting” refer to the control of bleeding, including thephysiological properties of vasoconstriction and coagulation. Bloodcoagulation assists in maintaining the integrity of mammaliancirculation after injury, inflammation, disease, congenital defect,dysfunction or other disruption. After initiation of clotting, bloodcoagulation proceeds through the sequential activation of certain plasmaproenzymes to their enzyme forms (see, for example, Coleman, R. W. etal. (eds.) Hemostasis and Thrombosis, Second Edition, (1987)). Theseplasma glycoproteins, including Factor XII, Factor XI, Factor IX, FactorX, Factor VII, and prothrombin, are zymogens of serine proteases. Mostof these blood clotting enzymes are effective on a physiological scaleonly when assembled in complexes on membrane surfaces with proteincofactors such as Factor VIII and Factor V. Other blood factors modulateand localize clot formation, or dissolve blood clots. Activated proteinC is a specific enzyme that inactivates procoagulant components. Calciumions are involved in many of the component reactions. Blood coagulationfollows either the intrinsic pathway, where all of the proteincomponents are present in blood, or the extrinsic pathway, where thecell-membrane protein tissue factor plays a critical role. Clotformation occurs when fibrinogen is cleaved by thrombin to form fibrin.Blood clots are composed of activated platelets and fibrin.

Further, the formation of blood clots does not only limit bleeding incase of an injury (hemostasis), but may lead to serious organ damage anddeath in the context of atherosclerotic diseases by occlusion of animportant artery or vein. Thrombosis is thus blood clot formation at thewrong time and place. It involves a cascade of complicated and regulatedbiochemical reactions between circulating blood proteins (coagulationfactors), blood cells (in particular platelets), and elements of aninjured vessel wall.

Accordingly, the present invention provides anticoagulation andantithrombotic treatments aiming at inhibiting the formation of bloodclots in order to prevent or treat blood coagulation disorders, such asmyocardial infarction, stroke, loss of a limb by peripheral arterydisease or pulmonary embolism.

As used interchangeably herein, “modulating or modulation of hemostasis”and “regulating or regulation of hemostasis” includes the induction(e.g., stimulation or increase) of hemostasis, as well as the inhibition(e.g., reduction or decrease) of hemostasis.

In one aspect, the invention provides a method for reducing orinhibiting hemostasis in a subject by administering a sirtuin-modulatingcompound that increases the level and/or activity of a sirtuin protein.The compositions and methods disclosed herein are useful for thetreatment or prevention of thrombotic disorders. As used herein, theterm “thrombotic disorder” includes any disorder or conditioncharacterized by excessive or unwanted coagulation or hemostaticactivity, or a hypercoagulable state. Thrombotic disorders includediseases or disorders involving platelet adhesion and thrombusformation, and may manifest as an increased propensity to formthromboses, e.g., an increased number of thromboses, thrombosis at anearly age, a familial tendency towards thrombosis, and thrombosis atunusual sites. Examples of thrombotic disorders include, but are notlimited to, thromboembolism, deep vein thrombosis, pulmonary embolism,stroke, myocardial infarction, miscarriage, thrombophilia associatedwith anti-thrombin III deficiency, protein C deficiency, protein Sdeficiency, resistance to activated protein C, dysfibrinogenemia,fibrinolytic disorders, homocystinuria, pregnancy, inflammatorydisorders, myeloproliferative disorders, arteriosclerosis, angina, e.g.,unstable angina, disseminated intravascular coagulation, thromboticthrombocytopenic purpura, cancer metastasis, sickle cell disease,glomerular nephritis, and drug induced thrombocytopenia (including, forexample, heparin induced thrombocytopenia). In addition,sirtuin-modulating compounds that increase the level and/or activity ofa sirtuin protein may be administered to prevent thrombotic events or toprevent re-occlusion during or after therapeutic clot lysis orprocedures such as angioplasty or surgery.

In another embodiment, a combination drug regimen may include drugs orcompounds for the treatment or prevention of blood coagulation disordersor secondary conditions associated with these conditions. Thus, acombination drug regimen may include one or more sirtuin-modulatingcompounds that increase the level and/or activity of a sirtuin proteinand one or more anti-coagulation or anti-thrombosis agents. For example,one or more sirtuin-modulating compounds can be combined with aneffective amount of one or more of: aspirin, heparin, and oral Warfarinthat inhibits Vit K-dependent factors, low molecular weight heparinsthat inhibit factors X and II, thrombin inhibitors, inhibitors ofplatelet GP IIbIIIa receptors, inhibitors of tissue factor (TF),inhibitors of human von Willebrand factor, inhibitors of one or morefactors involved in hemostasis (in particular in the coagulationcascade). In addition, sirtuin-modulating compounds that increase thelevel and/or activity of a sirtuin protein can be combined withthrombolytic agents, such as t-PA, streptokinase, reptilase, TNK-t-PA,and staphylokinase.

Weight Control

In another aspect, sirtuin-modulating compounds that increase the leveland/or activity of a sirtuin protein may be used for treating orpreventing weight gain or obesity in a subject. For example,sirtuin-modulating compounds that increase the level and/or activity ofa sirtuin protein may be used, for example, to treat or preventhereditary obesity, dietary obesity, hormone related obesity, obesityrelated to the administration of medication, to reduce the weight of asubject, or to reduce or prevent weight gain in a subject. A subject inneed of such a treatment may be a subject who is obese, likely to becomeobese, overweight, or likely to become overweight. Subjects who arelikely to become obese or overweight can be identified, for example,based on family history, genetics, diet, activity level, medicationintake, or various combinations thereof.

In yet other embodiments, sirtuin-modulating compounds that increase thelevel and/or activity of a sirtuin protein may be administered tosubjects suffering from a variety of other diseases and conditions thatmay be treated or prevented by promoting weight loss in the subject.Such diseases include, for example, high blood pressure, hypertension,high blood cholesterol, dyslipidemia, type 2 diabetes, insulinresistance, glucose intolerance, hyperinsulinemia, coronary heartdisease, angina pectoris, congestive heart failure, stroke, gallstones,cholescystitis and cholelithiasis, gout, osteoarritis, obstructive sleepapnea and respiratory problems, some types of cancer (such asendometrial, breast, prostate, and colon), complications of pregnancy,poor female reproductive health (such as menstrual irregularities,infertility, irregular ovulation), bladder control problems (such asstress incontinence); uric acid nephrolithiasis; psychological disorders(such as depression, eating disorders, distorted body image, and lowself esteem). Stunkard A J, Wadden T A. (Editors) Obesity: theory andtherapy, Second Edition. New York: Raven Press, 1993. Finally, patientswith AIDS can develop lipodystrophy or insulin resistance in response tocombination therapies for AIDS.

In another embodiment, sirtuin-modulating compounds that increase thelevel and/or activity of a sirtuin protein may be used for inhibitingadipogenesis or fat cell differentiation, whether in vitro or in vivo.In particular, high circulating levels of insulin and/or insulin likegrowth factor (IGF) 1 will be prevented from recruiting preadipocytes todifferentiate into adipocytes. Such methods may be used for treating orpreventing obesity.

In other embodiments, sirtuin-modulating compounds that increase thelevel and/or activity of a sirtuin protein may be used for reducingappetite and/or increasing satiety, thereby causing weight loss oravoidance of weight gain. A subject in need of such a treatment may be asubject who is overweight, obese or a subject likely to becomeoverweight or obese. The method may comprise administering daily or,every other day, or once a week, a dose, e.g., in the form of a pill, toa subject. The dose may be an “appetite reducing dose.”

In other embodiments, a sirtuin-modulating compound that decreases thelevel and/or activity of a sirtuin protein may be used to stimulateappetite and/or weight gain. A method may comprise administering to asubject, such as a subject in need thereof, a pharmaceutically effectiveamount of a sirtuin-modulating agent that decreases the level and/oractivity of a sirtuin protein, such as SIRT1 and/or SIRT3. A subject inneed of such a treatment may be a subject who has cachexia or may belikely to develop cachexia. A combination of agents may also beadministered. A method may further comprise monitoring in the subjectthe state of the disease or of activation of sirtuins, for example, inadipose tissue.

Methods for stimulating fat accumulation in cells may be used in vitro,to establish cell models of weight gain, which may be used, e.g., foridentifying other drugs that prevent weight gain.

Also provided are methods for modulating adipogenesis or fat celldifferentiation, whether in vitro or in vivo. In particular, highcirculating levels of insulin and/or insulin like growth factor (IGF) 1will be prevented from recruiting preadipocytes to differentiate intoadipocytes. Such methods may be used to modulate obesity. A method forstimulating adipogenesis may comprise contacting a cell with asirtuin-modulating agent that decreases the level and/or activity of asirtuin protein.

In another embodiment, the invention provides methods of decreasing fator lipid metabolism in a subject by administering a sirtuin-modulatingcompound that decreases the level and/or activity of a sirtuin protein.The method includes administering to a subject an amount of asirtuin-modulating compound, e.g., in an amount effective to decreasemobilization of fat to the blood from WAT cells and/or to decrease fatburning by BAT cells.

Methods for promoting appetite and/or weight gain may include, forexample, prior identifying a subject as being in need of decreased fator lipid metabolism, e.g., by weighing the subject, determining the BMIof the subject, or evaluating fat content of the subject or sirtuinactivity in cells of the subject. The method may also include monitoringthe subject, e.g., during and/or after administration of asirtuin-modulating compound. The administering can include one or moredosages, e.g., delivered in boluses or continuously. Monitoring caninclude evaluating a hormone or a metabolite. Exemplary hormones includeleptin, adiponectin, resistin, and insulin. Exemplary metabolitesinclude triglyercides, cholesterol, and fatty acids.

In one embodiment, a sirtuin-modulating compound that decreases thelevel and/or activity of a sirtuin protein may be used to modulate(e.g., increase) the amount of subcutaneous fat in a tissue, e.g., infacial tissue or in other surface-associated tissue of the neck, hand,leg, or lips. The sirtuin-modulating compound may be used to increasethe rigidity, water retention, or support properties of the tissue. Forexample, the sirtuin-modulating compound can be applied topically, e.g.,in association with another agent, e.g., for surface-associated tissuetreatment. The sirtuin-modulating compound may also be injectedsubcutaneously, e.g., within the region where an alteration insubcutaneous fat is desired.

A method for modulating weight may further comprise monitoring theweight of the subject and/or the level of modulation of sirtuins, forexample, in adipose tissue.

In an exemplary embodiment, sirtuin-modulating compounds that increasethe level and/or activity of a sirtuin protein may be administered as acombination therapy for treating or preventing weight gain or obesity.For example, one or more sirtuin-modulating compounds that increase thelevel and/or activity of a sirtuin protein may be administered incombination with one or more anti-obesity agents. Exemplary anti-obesityagents include, for example, phenylpropanolamine, ephedrine,pseudoephedrine, phentermine, a cholecystokinin-A agonist, a monoaminereuptake inhibitor (such as sibutramine), a sympathomimetic agent, aserotonergic agent (such as dexfenfluramine or fenfluramine), a dopamineagonist (such as bromocriptine), a melanocyte-stimulating hormonereceptor agonist or mimetic, a melanocyte-stimulating hormone analog, acannabinoid receptor antagonist, a melanin concentrating hormoneantagonist, the OB protein (leptin), a leptin analog, a leptin receptoragonist, a galanin antagonist or a GI lipase inhibitor or decreaser(such as orlistat). Other anorectic agents include bombesin agonists,dehydroepiandrosterone or analogs thereof, glucocorticoid receptoragonists and antagonists, orexin receptor antagonists, urocortin bindingprotein antagonists, agonists of the glucagon-like peptide-1 receptorsuch as Exendin and ciliary neurotrophic factors such as Axokine.

In another embodiment, sirtuin-modulating compounds that increase thelevel and/or activity of a sirtuin protein may be administered to reducedrug-induced weight gain. For example, a sirtuin-modulating compoundthat increases the level and/or activity of a sirtuin protein may beadministered as a combination therapy with medications that maystimulate appetite or cause weight gain, in particular, weight gain dueto factors other than water retention. Examples of medications that maycause weight gain, include for example, diabetes treatments, including,for example, sulfonylureas (such as glipizide and glyburide),thiazolidinediones (such as pioglitazone and rosiglitazone),meglitinides, nateglinide, repaglinide, sulphonylurea medicines, andinsulin; anti-depressants, including, for example, tricyclicantidepressants (such as amitriptyline and imipramine), irreversiblemonoamine oxidase inhibitors (MAOIs), selective serotonin reuptakeinhibitors (SSRIs), bupropion, paroxetine, and mirtazapine; steroids,such as, for example, prednisone; hormone therapy; lithium carbonate;valproic acid; carbamazepine; chlorpromazine; thiothixene; beta blockers(such as propranolo); alpha blockers (such as clonidine, prazosin andterazosin); and contraceptives including oral contraceptives (birthcontrol pills) or other contraceptives containing estrogen and/orprogesterone (Depo-Provera, Norplant, Ortho), testosterone or Megestrol.In another exemplary embodiment, sirtuin-modulating compounds thatincrease the level and/or activity of a sirtuin protein may beadministered as part of a smoking cessation program to prevent weightgain or reduce weight already gained.

Metabolic Disorders/Diabetes

In another aspect, sirtuin-modulating compounds that increase the leveland/or activity of a sirtuin protein may be used for treating orpreventing a metabolic disorder, such as insulin-resistance, apre-diabetic state, type II diabetes, and/or complications thereof.Administration of a sirtuin-modulating compounds that increases thelevel and/or activity of a sirtuin protein may increase insulinsensitivity and/or decrease insulin levels in a subject. A subject inneed of such a treatment may be a subject who has insulin resistance orother precursor symptom of type II diabetes, who has type II diabetes,or who is likely to develop any of these conditions. For example, thesubject may be a subject having insulin resistance, e.g., having highcirculating levels of insulin and/or associated conditions, such ashyperlipidemia, dyslipogenesis, hypercholesterolemia, impaired glucosetolerance, high blood glucose sugar level, other manifestations ofsyndrome X, hypertension, atherosclerosis and lipodystrophy.

In an exemplary embodiment, sirtuin-modulating compounds that increasethe level and/or activity of a sirtuin protein may be administered as acombination therapy for treating or preventing a metabolic disorder. Forexample, one or more sirtuin-modulating compounds that increase thelevel and/or activity of a sirtuin protein may be administered incombination with one or more anti-diabetic agents. Exemplaryanti-diabetic agents include, for example, an aldose reductaseinhibitor, a glycogen phosphorylase inhibitor, a sorbitol dehydrogenaseinhibitor, a protein tyrosine phosphatase 1B inhibitor, a dipeptidylprotease inhibitor, insulin (including orally bioavailable insulinpreparations), an insulin mimetic, metformin, acarbose, a peroxisomeproliferator-activated receptor-γ (PPAR-γ) ligand such as troglitazone,rosaglitazone, pioglitazone or GW-1929, a sulfonylurea, glipazide,glyburide, or chlorpropamide wherein the amounts of the first and secondcompounds result in a therapeutic effect. Other anti-diabetic agentsinclude a glucosidase inhibitor, a glucagon-like peptide-1 (GLP-1),insulin, a PPAR α/γ dual agonist, a meglitimide and an aP2 inhibitor. Inan exemplary embodiment, an anti-diabetic agent may be a dipeptidylpeptidase IV (DP-IV or DPP-IV) inhibitor, such as, for example LAF237from Novartis (NVP DPP728;1-[[[2-[(5-cyanopyridin-2-yl)amino]ethyl]amino]acetyl]-2-cyano-(S)-pyrrolidine)or MK-04301 from Merck (see e.g., Hughes et al., Biochemistry 38:11597-603 (1999)).

Inflammatory Diseases

In other aspects, sirtuin-modulating compounds that increase the leveland/or activity of a sirtuin protein can be used to treat or prevent adisease or disorder associated with inflammation. Sirtuin-modulatingcompounds that increase the level and/or activity of a sirtuin proteinmay be administered prior to the onset of, at, or after the initiationof inflammation. When used prophylactically, the compounds arepreferably provided in advance of any inflammatory response or symptom.Administration of the compounds may prevent or attenuate inflammatoryresponses or symptoms.

Exemplary inflammatory conditions include, for example, multiplesclerosis, rheumatoid arthritis, psoriatic arthritis, degenerative jointdisease, spondouloarthropathies, gouty arthritis, systemic lupuserythematosus, juvenile arthritis, rheumatoid arthritis, osteoarthritis,osteoporosis, diabetes (e.g., insulin dependent diabetes mellitus orjuvenile onset diabetes), menstrual cramps, cystic fibrosis,inflammatory bowel disease, irritable bowel syndrome, Crohn's disease,mucous colitis, ulcerative colitis, gastritis, esophagitis,pancreatitis, peritonitis, Alzheimer's disease, shock, ankylosingspondylitis, gastritis, conjunctivitis, pancreatis (acute or chronic),multiple organ injury syndrome (e.g., secondary to septicemia ortrauma), myocardial infarction, atherosclerosis, stroke, reperfusioninjury (e.g., due to cardiopulmonary bypass or kidney dialysis), acuteglomerulonephritis, vasculitis, thermal injury (i.e., sunburn),necrotizing enterocolitis, granulocyte transfusion associated syndrome,and/or Sjogren's syndrome. Exemplary inflammatory conditions of the skininclude, for example, eczema, atopic dermatitis, contact dermatitis,urticaria, schleroderma, psoriasis, and dermatosis with acuteinflammatory components.

In another embodiment, sirtuin-modulating compounds that increase thelevel and/or activity of a sirtuin protein may be used to treat orprevent allergies and respiratory conditions, including asthma,bronchitis, pulmonary fibrosis, allergic rhinitis, oxygen toxicity,emphysema, chronic bronchitis, acute respiratory distress syndrome, andany chronic obstructive pulmonary disease (COPD). The compounds may beused to treat chronic hepatitis infection, including hepatitis B andhepatitis C.

Additionally, sirtuin-modulating compounds that increase the leveland/or activity of a sirtuin protein may be used to treat autoimmunediseases and/or inflammation associated with autoimmune diseases such asorgan-tissue autoimmune diseases (e.g., Raynaud's syndrome),scleroderma, myasthenia gravis, transplant rejection, endotoxin shock,sepsis, psoriasis, eczema, dermatitis, multiple sclerosis, autoimmunethyroiditis, uveitis, systemic lupus erythematosis, Addison's disease,autoimmune polyglandular disease (also known as autoimmune polyglandularsyndrome), and Grave's disease.

In certain embodiments, one or more sirtuin-modulating compounds thatincrease the level and/or activity of a sirtuin protein may be takenalone or in combination with other compounds useful for treating orpreventing inflammation. Exemplary anti-inflammatory agents include, forexample, steroids (e.g., cortisol, cortisone, fludrocortisone,prednisone, 6α-methylprednisone, triamcinolone, betamethasone ordexamethasone), nonsteroidal antiinflammatory drugs (NSAIDS (e.g.,aspirin, acetaminophen, tolmetin, ibuprofen, mefenamic acid, piroxicam,nabumetone, rofecoxib, celecoxib, etodolac or nimesulide). In anotherembodiment, the other therapeutic agent is an antibiotic (e.g.,vancomycin, penicillin, amoxicillin, ampicillin, cefotaxime,ceftriaxone, cefixime, rifampinmetronidazole, doxycycline orstreptomycin). In another embodiment, the other therapeutic agent is aPDE4 inhibitor (e.g., roflumilast or rolipram). In another embodiment,the other therapeutic agent is an antihistamine (e.g., cyclizine,hydroxyzine, promethazine or diphenhydramine). In another embodiment,the other therapeutic agent is an anti-malarial (e.g., artemisinin,artemether, artsunate, chloroquine phosphate, mefloquine hydrochloride,doxycycline hyclate, proguanil hydrochloride, atovaquone orhalofantrine). In one embodiment, the other therapeutic agent isdrotrecogin alfa.

Further examples of anti-inflammatory agents include, for example,aceclofenac, acemetacin, e-acetamidocaproic acid, acetaminophen,acetaminosalol, acetanilide, acetylsalicylic acid, S-adenosylmethionine,alclofenac, alclometasone, alfentanil, algestone, allylprodine,alminoprofen, aloxiprin, alphaprodine, aluminum bis(acetylsalicylate),amcinonide, amfenac, aminochlorthenoxazin, 3-amino-4-hydroxybutyricacid, 2-amino-4-picoline, aminopropylon, aminopyrine, amixetrine,ammonium salicylate, ampiroxicam, amtolmetin guacil, anileridine,antipyrine, antrafenine, apazone, beclomethasone, bendazac, benorylate,benoxaprofen, benzpiperylon, benzydamine, benzylmorphine, bermoprofen,betamethasone, betamethasone-17-valerate, bezitramide, α-bisabolol,bromfenac, p-bromoacetanilide, 5-bromosalicylic acid acetate,bromosaligenin, bucetin, bucloxic acid, bucolome, budesonide, bufexamac,bumadizon, buprenorphine, butacetin, butibufen, butorphanol,carbamazepine, carbiphene, carprofen, carsalam, chlorobutanol,chloroprednisone, chlorthenoxazin, choline salicylate, cinchophen,cinmetacin, ciramadol, clidanac, clobetasol, clocortolone, clometacin,clonitazene, clonixin, clopirac, cloprednol, clove, codeine, codeinemethyl bromide, codeine phosphate, codeine sulfate, cortisone,cortivazol, cropropamide, crotethamide, cyclazocine, deflazacort,dehydrotestosterone, desomorphine, desonide, desoximetasone,dexamethasone, dexamethasone-21-isonicotinate, dexoxadrol,dextromoramide, dextropropoxyphene, deoxycorticosterone, dezocine,diampromide, diamorphone, diclofenac, difenamizole, difenpiramide,diflorasone, diflucortolone, diflunisal, difluprednate, dihydrocodeine,dihydrocodeinone enol acetate, dihydromorphine, dihydroxyaluminumacetylsalicylate, dimenoxadol, dimepheptanol, dimethylthiambutene,dioxaphetyl butyrate, dipipanone, diprocetyl, dipyrone, ditazol,droxicam, emorfazone, enfenamic acid, enoxolone, epirizole, eptazocine,etersalate, ethenzamide, ethoheptazine, ethoxazene,ethylmethylthiambutene, ethylmorphine, etodolac, etofenamate,etonitazene, eugenol, felbinac, fenbufen, fenclozic acid, fendosal,fenoprofen, fentanyl, fentiazac, fepradinol, feprazone, floctafenine,fluazacort, flucloronide, flufenamic acid, flumethasone, flunisolide,flunixin, flunoxaprofen, fluocinolone acetonide, fluocinonide,fluocinolone acetonide, fluocortin butyl, fluocortolone, fluoresone,fluorometholone, fluperolone, flupirtine, fluprednidene,fluprednisolone, fluproquazone, flurandrenolide, flurbiprofen,fluticasone, formocortal, fosfosal, gentisic acid, glafenine,glucametacin, glycol salicylate, guaiazulene, halcinonide, halobetasol,halometasone, haloprednone, heroin, hydrocodone, hydrocortamate,hydrocortisone, hydrocortisone acetate, hydrocortisone succinate,hydrocortisone hemisuccinate, hydrocortisone 21-lysinate, hydrocortisonecypionate, hydromorphone, hydroxypethidine, ibufenac, ibuprofen,ibuproxam, imidazole salicylate, indomethacin, indoprofen, isofezolac,isoflupredone, isoflupredone acetate, isoladol, isomethadone, isonixin,isoxepac, isoxicam, ketobemidone, ketoprofen, ketorolac,p-lactophenetide, lefetamine, levallorphan, levorphanol,levophenacyl-morphan, lofentanil, lonazolac, lornoxicam, loxoprofen,lysine acetylsalicylate, mazipredone, meclofenamic acid, medrysone,mefenamic acid, meloxicam, meperidine, meprednisone, meptazinol,mesalamine, metazocine, methadone, methotrimeprazine,methylprednisolone, methylprednisolone acetate, methylprednisolonesodium succinate, methylprednisolone suleptnate, metiazinic acid,metofoline, metopon, mofebutazone, mofezolac, mometasone, morazone,morphine, morphine hydrochloride, morphine sulfate, morpholinesalicylate, myrophine, nabumetone, nalbuphine, nalorphine, 1-naphthylsalicylate, naproxen, narceine, nefopam, nicomorphine, nifenazone,niflumic acid, nimesulide, 5′-nitro-2′-propoxyacetanilide,norlevorphanol, normethadone, normorphine, norpipanone, olsalazine,opium, oxaceprol, oxametacine, oxaprozin, oxycodone, oxymorphone,oxyphenbutazone, papavereturn, paramethasone, paranyline, parsalmide,pentazocine, perisoxal, phenacetin, phenadoxone, phenazocine,phenazopyridine hydrochloride, phenocoll, phenoperidine, phenopyrazone,phenomorphan, phenyl acetylsalicylate, phenylbutazone, phenylsalicylate, phenyramidol, piketoprofen, piminodine, pipebuzone,piperylone, pirazolac, piritramide, piroxicam, pirprofen, pranoprofen,prednicarbate, prednisolone, prednisone, prednival, prednylidene,proglumetacin, proheptazine, promedol, propacetamol, properidine,propiram, propoxyphene, propyphenazone, proquazone, protizinic acid,proxazole, ramifenazone, remifentanil, rimazolium metilsulfate,salacetamide, salicin, salicylamide, salicylamide o-acetic acid,salicylic acid, salicylsulfuric acid, salsalate, salverine, simetride,sufentanil, sulfasalazine, sulindac, superoxide dismutase, suprofen,suxibuzone, talniflumate, tenidap, tenoxicam, terofenamate, tetrandrine,thiazolinobutazone, tiaprofenic acid, tiaramide, tilidine, tinoridine,tixocortol, tolfenamic acid, tolmetin, tramadol, triamcinolone,triamcinolone acetonide, tropesin, viminol, xenbucin, ximoprofen,zaltoprofen and zomepirac.

In an exemplary embodiment, a sirtuin-modulating compound that increasesthe level and/or activity of a sirtuin protein may be administered witha selective COX-2 inhibitor for treating or preventing inflammation.Exemplary selective COX-2 inhibitors include, for example, deracoxib,parecoxib, celecoxib, valdecoxib, rofecoxib, etoricoxib, lumiracoxib,2-(3,5-difluorophenyl)-3-[4-(methylsulfonyl)phenyl]-2-cyclopenten-1-one,(S)-6,8-dichloro-2-(triflu- oromethyl)-2H-1-benzopyran-3-carboxylicacid,2-(3,4-difluorophenyl)-4-(3-hydroxy-3-methyl-1-butoxy)-5-[4-(methylsulfonyl)phenyl]-3-(2H)-pyridazinone,4-[5-(4-fluorophenyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl]benzenesulfonamide,tert-butyl 1benzyl-4-[(4-oxopiperidin-1-yl}sulfonyl]piperidine-4-carboxylate,4-[5-(phenyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl]benzenesulfonamide,salts and prodrugs thereof.

Flushing

In another aspect, sirtuin-modulating compounds that increase the leveland/or activity of a sirtuin protein may be used for reducing theincidence or severity of flushing and/or hot flashes which are symptomsof a disorder. For instance, the subject method includes the use ofsirtuin-modulating compounds that increase the level and/or activity ofa sirtuin protein, alone or in combination with other agents, forreducing incidence or severity of flushing and/or hot flashes in cancerpatients. In other embodiments, the method provides for the use ofsirtuin-modulating compounds that increase the level and/or activity ofa sirtuin protein to reduce the incidence or severity of flushing and/orhot flashes in menopausal and post-menopausal woman.

In another aspect, sirtuin-modulating compounds that increase the leveland/or activity of a sirtuin protein may be used as a therapy forreducing the incidence or severity of flushing and/or hot flashes whichare side-effects of another drug therapy, e.g., drug-induced flushing.In certain embodiments, a method for treating and/or preventingdrug-induced flushing comprises administering to a patient in needthereof a formulation comprising at least one flushing inducing compoundand at least one sirtuin-modulating compound that increases the leveland/or activity of a sirtuin protein. In other embodiments, a method fortreating drug induced flushing comprises separately administering one ormore compounds that induce flushing and one or more sirtuin-modulatingcompounds, e.g., wherein the sirtuin-modulating compound and flushinginducing agent have not been formulated in the same compositions. Whenusing separate formulations, the sirtuin-modulating compound may beadministered (1) at the same as administration of the flushing inducingagent, (2) intermittently with the flushing inducing agent, (3)staggered relative to administration of the flushing inducing agent, (4)prior to administration of the flushing inducing agent, (5) subsequentto administration of the flushing inducing agent, and (6) variouscombination thereof. Exemplary flushing inducing agents include, forexample, niacin, faloxifene, antidepressants, anti-psychotics,chemotherapeutics, calcium channel blockers, and antibiotics.

In one embodiment, sirtuin-modulating compounds that increase the leveland/or activity of a sirtuin protein may be used to reduce flushing sideeffects of a vasodilator or an antilipemic agent (includinganticholesteremic agents and lipotropic agents). In an exemplaryembodiment, a sirtuin-modulating compound that increases the leveland/or activity of a sirtuin protein may be used to reduce flushingassociated with the administration of niacin.

Nicotinic acid, 3-pyridinecarboxylic acid or niacin, is an antilipidemicagent that is marketed under, for example, the trade names Nicolar®,SloNiacin®, Nicobid® and Time Release Niacin®. Nicotinic acid has beenused for many years in the treatment of lipidemic disorders such ashyperlipidemia, hypercholesterolemia and atherosclerosis. This compoundhas long been known to exhibit the beneficial effects of reducing totalcholesterol, low density lipoproteins or “LDL cholesterol,”triglycerides and apolipoprotein a (Lp(a)) in the human body, whileincreasing desirable high density lipoproteins or “HDL cholesterol”.

Typical doses range from about 1 gram to about 3 grams daily. Nicotinicacid is normally administered two to four times per day after meals,depending upon the dosage form selected. Nicotinic acid is currentlycommercially available in two dosage forms. One dosage form is animmediate or rapid release tablet which should be administered three orfour times per day. Immediate release (“IR”) nicotinic acid formulationsgenerally release nearly all of their nicotinic acid within about 30 to60 minutes following ingestion. The other dosage form is a sustainedrelease form which is suitable for administration two to four times perday. In contrast to IR formulations, sustained release (“SR”) nicotinicacid formulations are designed to release significant quantities of drugfor absorption into the blood stream over specific timed intervals inorder to maintain therapeutic levels of nicotinic acid over an extendedperiod such as 12 or 24 hours after ingestion.

As used herein, the term “nicotinic acid” is meant to encompassnicotinic acid or a compound other than nicotinic acid itself which thebody metabolizes into nicotinic acid, thus producing essentially thesame effect as nicotinic acid. Exemplary compounds that produce aneffect similar to that of nicotinic acid include, for example, nicotinylalcohol tartrate, d-glucitol hexanicotinate, aluminum nicotinate,niceritrol and d, 1-alpha-tocopheryl nicotinate. Each such compound willbe collectively referred to herein as “nicotinic acid.”

In another embodiment, the invention provides a method for treatingand/or preventing hyperlipidemia with reduced flushing side effects. Themethod comprises the steps of administering to a subject in need thereofa therapeutically effective amount of nicotinic acid and asirtuin-modulating compound that increases the level and/or activity ofa sirtuin protein in an amount sufficient to reduce flushing. In anexemplary embodiment, the nicotinic acid and/or sirtuin-modulatingcompound may be administered nocturnally.

In another representative embodiment, the method involves the use ofsirtuin-modulating compounds that increase the level and/or activity ofa sirtuin protein to reduce flushing side effects of raloxifene.Raloxifene acts like estrogen in certain places in the body, but is nota hormone. It helps prevent osteoporosis in women who have reachedmenopause. Osteoporosis causes bones to gradually grow thin, fragile,and more likely to break. Evista slows down the loss of bone mass thatoccurs with menopause, lowering the risk of spine fractures due toosteoporosis. A common side effect of raloxifene is hot flashes(sweating and flushing). This can be uncomfortable for women who alreadyhave hot flashes due to menopause.

In another representative embodiment, the method involves the use ofsirtuin-modulating compounds that increase the level and/or activity ofa sirtuin protein to reduce flushing side effects of antidepressants oranti-psychotic agent. For instance, sirtuin-modulating compounds thatincrease the level and/or activity of a sirtuin protein can be used inconjunction (administered separately or together) with a serotoninreuptake inhibitor, a 5HT2 receptor antagonist, an anticonvulsant, anorepinephrine reuptake inhibitor, an α-adrenoreceptor antagonist, anNK-3 antagonist, an NK-1 receptor antagonist, a PDE4 inhibitor, anNeuropeptide Y5 Receptor Antagonists, a D4 receptor antagonist, a 5HT1Areceptor antagonist, a 5HT1D receptor antagonist, a CRF antagonist, amonoamine oxidase inhibitor, or a sedative-hypnotic drug.

In certain embodiments, sirtuin-modulating compounds that increase thelevel and/or activity of a sirtuin protein may be used as part of atreatment with a serotonin reuptake inhibitor (SRI) to reduce flushing.In certain preferred embodiments, the SRI is a selective serotoninreuptake inhibitor (SSRI), such as a fluoxetinoid (fluoxetine,norfluoxetine) or a nefazodonoid (nefazodone, hydroxynefazodone,oxonefazodone). Other exemplary SSRI's include duloxetine, venlafaxine,milnacipran, citalopram, fluvoxamine, paroxetine and sertraline. Thesirtuin-modulating compound that increases the level and/or activity ofa sirtuin protein can also be used as part of a treatment withsedative-hypnotic drug, such as selected from the group consisting of abenzodiazepine (such as alprazolam, chlordiazepoxide, clonazepam,chlorazepate, clobazam, diazepam, halazepam, lorazepam, oxazepam andprazepam), zolpidem, and barbiturates. In still other embodiments, asirtuin-modulating compound that increases the level and/or activity ofa sirtuin protein may be used as part of a treatment with a 5-HT1Areceptor partial agonist, such as selected from the group consisting ofbuspirone, flesinoxan, gepirone and ipsapirone. Sirtuin-modulatingcompounds that increase the level and/or activity of a sirtuin proteincan also used as part of a treatment with a norepinephrine reuptakeinhibitor, such as selected from tertiary amine tricyclics and secondaryamine tricyclics. Exemplary tertiary amine tricyclic includeamitriptyline, clomipramine, doxepin, imipramine and trimipramine.Exemplary secondary amine tricyclic include amoxapine, desipramine,maprotiline, nortriptyline and protriptyline. In certain embodiments,sirtuin-modulating compounds that increase the level and/or activity ofa sirtuin protein may be used as part of a treatment with a monoamineoxidase inhibitor, such as selected from the group consisting ofisocarboxazid, phenelzine, tranylcypromine, selegiline and moclobemide.

In still another representative embodiment, sirtuin-modulating compoundsthat increase the level and/or activity of a sirtuin protein may be usedto reduce flushing side effects of chemotherapeutic agents, such ascyclophosphamide, tamoxifen.

In another embodiment, sirtuin-modulating compounds that increase thelevel and/or activity of a sirtuin protein may be used to reduceflushing side effects of calcium channel blockers, such as amlodipine.

In another embodiment, sirtuin-modulating compounds that increase thelevel and/or activity of a sirtuin protein may be used to reduceflushing side effects of antibiotics. For example, sirtuin-modulatingcompounds that increase the level and/or activity of a sirtuin proteincan be used in combination with levofloxacin. Levofloxacin is used totreat infections of the sinuses, skin, lungs, ears, airways, bones, andjoints caused by susceptible bacteria. Levofloxacin also is frequentlyused to treat urinary infections, including those resistant to otherantibiotics, as well as prostatitis. Levofloxacin is effective intreating infectious diarrheas caused by E. coli, campylobacter jejuni,and shigella bacteria. Levofloxacin also can be used to treat variousobstetric infections, including mastitis.

Ocular Disorders

One aspect of the present invention is a method for inhibiting, reducingor otherwise treating vision impairment by administering to a patient atherapeutic dosage of sirtuin modulator selected from a compounddisclosed herein, or a pharmaceutically acceptable salt, prodrug or ametabolic derivative thereof.

In certain aspects of the invention, the vision impairment is caused bydamage to the optic nerve or central nervous system. In particularembodiments, optic nerve damage is caused by high intraocular pressure,such as that created by glaucoma. In other particular embodiments, opticnerve damage is caused by swelling of the nerve, which is oftenassociated with an infection or an immune (e.g., autoimmune) responsesuch as in optic neuritis.

Glaucoma describes a group of disorders which are associated with avisual field defect, cupping of the optic disc, and optic nerve damage.These are commonly referred to as glaucomatous optic neuropathies. Mostglaucomas are usually, but not always, associated with a rise inintraocular pressure. Exemplary forms of glaucoma include Glaucoma andPenetrating Keratoplasty, Acute Angle Closure, Chronic Angle Closure,Chronic Open Angle, Angle Recession, Aphakic and Pseudophakic,Drug-Induced, Hyphema, Intraocular Tumors, Juvenile, Lens-Particle, LowTension, Malignant, Neovascular, Phacolytic, Phacomorphic, Pigmentary,Plateau Iris, Primary Congenital, Primary Open Angle, Pseudoexfoliation,Secondary Congenital, Adult Suspect, Unilateral, Uveitic, OcularHypertension, Ocular Hypotony, Posner-Schlossman Syndrome and ScleralExpansion Procedure in Ocular Hypertension & Primary Open-angleGlaucoma.

Intraocular pressure can also be increased by various surgicalprocedures, such as phacoemulsification (i.e., cataract surgery) andimplanation of structures such as an artificial lens. In addition,spinal surgeries in particular, or any surgery in which the patient isprone for an extended period of time can lead to increased interoccularpressure.

Optic neuritis (ON) is inflammation of the optic nerve and causes acuteloss of vision. It is highly associated with multiple sclerosis (MS) as15-25% of MS patients initially present with ON, and 50-75% of ONpatients are diagnosed with MS. ON is also associated with infection(e.g., viral infection, meningitis, syphilis), inflammation (e.g., froma vaccine), infiltration and ischemia.

Another condition leading to optic nerve damage is anterior ischemicoptic neuropathy (AION). There are two types of AION. Arteritic AION isdue to giant cell arteritis (vasculitis) and leads to acute vision loss.Non-arteritic AION encompasses all cases of ischemic optic neuropathyother than those due to giant cell arteritis. The pathophysiology ofAION is unclear although it appears to incorporate both inflammatory andischemic mechanisms.

Other damage to the optic nerve is typically associated withdemyleination, inflammation, ischemia, toxins, or trauma to the opticnerve. Exemplary conditions where the optic nerve is damaged includeDemyelinating Optic Neuropathy (Optic Neuritis, Retrobulbar OpticNeuritis), Optic Nerve Sheath Meningioma, Adult Optic Neuritis,Childhood Optic Neuritis, Anterior Ischemic Optic Neuropathy, PosteriorIschemic Optic Neuropathy, Compressive Optic Neuropathy, Papilledema,Pseudopapilledema and Toxic/Nutritional Optic Neuropathy.

Other neurological conditions associated with vision loss, albeit notdirectly associated with damage to the optic nerve, include Amblyopia,Bells Palsy, Chronic Progressive External Opthalmoplegia, MultipleSclerosis, Pseudotumor Cerebri and Trigeminal Neuralgia.

In certain aspects of the invention, the vision impairment is caused byretinal damage. In particular embodiments, retinal damage is caused bydisturbances in blood flow to the eye (e.g., arteriosclerosis,vasculitis). In particular embodiments, retinal damage is caused bydisrupton of the macula (e.g., exudative or non-exudative maculardegeneration).

Exemplary retinal diseases include Exudative Age Related MacularDegeneration, Nonexudative Age Related Macular Degeneration, RetinalElectronic Prosthesis and RPE Transplantation Age Related MacularDegeneration, Acute Multifocal Placoid Pigment Epitheliopathy, AcuteRetinal Necrosis, Best Disease, Branch Retinal Artery Occlusion, BranchRetinal Vein Occlusion, Cancer Associated and Related AutoimmuneRetinopathies, Central Retinal Artery Occlusion, Central Retinal VeinOcclusion, Central Serous Chorioretinopathy, Eales Disease, EpimacularMembrane, Lattice Degeneration, Macroaneurysm, Diabetic Macular Edema,Irvine-Gass Macular Edema, Macular Hole, Subretinal NeovascularMembranes, Diffuse Unilateral Subacute Neuroretinitis, NonpseudophakicCystoid Macular Edema, Presumed Ocular Histoplasmosis Syndrome,Exudative Retinal Detachment, Postoperative Retinal Detachment,Proliferative Retinal Detachment, Rhegmatogenous Retinal Detachment,Tractional Retinal Detachment, Retinitis Pigmentosa, CMV Retinitis,Retinoblastoma, Retinopathy of Prematurity, Birdshot Retinopathy,Background Diabetic Retinopathy, Proliferative Diabetic Retinopathy,Hemoglobinopathies Retinopathy, Purtscher Retinopathy, ValsalvaRetinopathy, Juvenile Retinoschisis, Senile Retinoschisis, TersonSyndrome and White Dot Syndromes.

Other exemplary diseases include ocular bacterial infections (e.g.conjunctivitis, keratitis, tuberculosis, syphilis, gonorrhea), viralinfections (e.g. Ocular Herpes Simplex Virus, Varicella Zoster Virus,Cytomegalovirus retinitis, Human Immunodeficiency Virus (HIV)) as wellas progressive outer retinal necrosis secondary to HIV or otherHIV-associated and other immunodeficiency-associated ocular diseases. Inaddition, ocular diseases include fungal infections (e.g. Candidachoroiditis, histoplasmosis), protozoal infections (e.g. toxoplasmosis)and others such as ocular toxocariasis and sarcoidosis.

One aspect of the invention is a method for inhibiting, reducing ortreating vision impairment in a subject undergoing treatment with achemotherapeutic drug (e.g., a neurotoxic drug, a drug that raisesintraocular pressure such as a steroid), by administering to the subjectin need of such treatment a therapeutic dosage of a sirtuin modulatordisclosed herein.

Another aspect of the invention is a method for inhibiting, reducing ortreating vision impairment in a subject undergoing surgery, includingocular or other surgeries performed in the prone position such as spinalcord surgery, by administering to the subject in need of such treatmenta therapeutic dosage of a sirtuin modulator disclosed herein. Ocularsurgeries include cataract, iridotomy and lens replacements. Anotheraspect of the invention is the treatment, including inhibition andprophylactic treatment, of age related ocular diseases includecataracts, dry eye, retinal damage and the like, by administering to thesubject in need of such treatment a therapeutic dosage of a sirtuinmodulator disclosed herein.

The formation of cataracts is associated with several biochemicalchanges in the lens of the eye, such as decreased levels of antioxidantsascorbic acid and glutathione, increased lipid, amino acid and proteinoxidation, increased sodium and calcium, loss of amino acids anddecreased lens metabolism. The lens, which lacks blood vessels, issuspended in extracellular fluids in the anterior part of the eye.Nutrients, such as ascorbic acid, glutathione, vitamin E, selenium,bioflavonoids and carotenoids are required to maintain the transparencyof the lens. Low levels of selenium results in an increase of freeradical-inducing hydrogen peroxide, which is neutralized by theselenium-dependent antioxidant enzyme glutathione peroxidase.Lens-protective glutathione peroxidase is also dependent on the aminoacids methionine, cysteine, glycine and glutamic acid.

Cataracts can also develop due to an inability to properly metabolizegalactose found in dairy products that contain lactose, a disaccharidecomposed of the monosaccharide galactose and glucose. Cataracts can beprevented, delayed, slowed and possibly even reversed if detected earlyand metabolically corrected.

Retinal damage is attributed, inter alia, to free radical initiatedreactions in glaucoma, diabetic tetinopathy and age-related maculardegeneration (AMD). The eye is a part of the central nervous system andhas limited regenerative capability. The retina is composed of numerousnerve cells which contain the highest concentration of polyunsaturatedfatty acids (PFA) and subject to oxidation. Free radicals are generatedby UV light entering the eye and mitochondria in the rods and cones,which generate the energy necessary to transform light into visualimpulses. Free radicals cause peroxidation of the PFA by hydroxyl orsuperoxide radicals which in turn propagate additional free radicals.The free radicals cause temporary or permanent damage to retinal tissue.

Glaucoma is usually viewed as a disorder that causes an elevatedintraocular pressure (IOP) that results in permanent damage to theretinal nerve fibers, but a sixth of all glaucoma cases do not developan elevated IOP. This disorder is now perceived as one of reducedvascular perfusion and an increase in neurotoxic factors. Recent studieshave implicated elevated levels of glutamate, nitric oxide andperoxynitirite in the eye as the causes of the death of retinal ganglioncells. Neuroprotective agents may be the future of glaucoma care. Forexample, nitric oxide synthase inhibitors block the formation ofperoxynitrite from nitric oxide and superoxide. In a recent study,animals treated with aminoguanidine, a nitric oxide synthase inhibitor,had a reduction in the loss of retinal ganglion cells. It was concludedthat nitric oxide in the eye caused cytotoxicity in many tissues andneurotoxicity in the central nervous system.

Diabetic retinopathy occurs when the underlying blood vessels developmicrovascular abnormalities consisting primarily of microaneurysms andintraretinal hemorrhages. Oxidative metabolites are directly involvedwith the pathogenesis of diabetic retinopathy and free radicals augmentthe generation of growth factors that lead to enhanced proliferativeactivity. Nitric oxide produced by endothelial cells of the vessels mayalso cause smooth muscle cells to relax and result in vasodilation ofsegments of the vessel. Ischemia and hypoxia of the retina occur afterthickening of the arterial basement membrane, endothelial proliferationand loss of pericytes. The inadequate oxygenation causes capillaryobliteration or nonperfusion, arteriolar-venular shunts, sluggish bloodflow and an impaired ability of RBCs to release oxygen. Lipidperoxidation of the retinal tissues also occurs as a result of freeradical damage.

The macula is responsible for our acute central vision and composed oflight-sensing cells (cones) while the underlying retinal pigmentepithelium (RPE) and choroid nourish and help remove waste materials.The RPE nourishes the cones with the vitamin A substrate for thephotosensitive pigments and digests the cones shed outer tips. RPE isexposed to high levels of UV radiation, and secretes factors thatinhibit angiogenesis. The choroid contains a dense vascular network thatprovides nutrients and removes the waste materials.

In AMD, the shed cone tips become indigestible by the RPE, where thecells swell and die after collecting too much undigested material.Collections of undigested waste material, called drusen, form under theRPE. Photoxic damage also causes the accumulation of lipofuscin in RPEcells. The intracellular lipofuscin and accumulation of drusen inBruch's membrane interferes with the transport of oxygen and nutrientsto the retinal tissues, and ultimately leads to RPE and photoreceptordysfunction. In exudative AMD, blood vessels grow from thechoriocapillaris through defects in Bruch's membrane and may grow underthe RPE, detaching it from the choroid, and leaking fluid or bleeding.

Macular pigment, one of the protective factors that prevent sunlightfrom damaging the retina, is formed by the accumulation of nutritionallyderived carotenoids, such as lutein, the fatty yellow pigment thatserves as a delivery vehicle for other important nutrients andzeaxanthin. Antioxidants such as vitamins C and E, beta-carotene andlutein, as well as zinc, selenium and copper, are all found in thehealthy macula. In addition to providing nourishment, these antioxidantsprotect against free radical damage that initiates macular degeneration.

Another aspect of the invention is the prevention or treatment of damageto the eye caused by stress, chemical insult or radiation, byadministering to the subject in need of such treatment a therapeuticdosage of a sirtuin modulator disclosed herein. Radiation orelectromagnetic damage to the eye can include that caused by CRT's orexposure to sunlight or UV.

In one embodiment, a combination drug regimen may include drugs orcompounds for the treatment or prevention of ocular disorders orsecondary conditions associated with these conditions. Thus, acombination drug regimen may include one or more sirtuin activators andone or more therapeutic agents for the treatment of an ocular disorder.For example, one or more sirtuin-activating compounds can be combinedwith an effective amount of one or more of: an agent that reducesintraocular pressure, an agent for treating glaucoma, an agent fortreating optic neuritis, an agent for treating CMV Retinopathy, an agentfor treating multiple sclerosis, and/or an antibiotic, etc.

In one embodiment, a sirtuin modulator can be administered inconjunction with a therapy for reducing intraocular pressure. One groupof therapies involves blocking aqueous production. For example, topicalbeta-adrenergic antagonists (timolol and betaxolol) decrease aqueousproduction. Topical timolol causes IOP to fall in 30 minutes with peakeffects in 1-2 hours. A reasonable regimen is Timoptic 0.5%, one dropevery 30 minutes for 2 doses. The carbonic anhydrase inhibitor,acetazolamide, also decreases aqueous production and should be given inconjunction with topical beta-antagonists. An initial dose of 500 mg isadministered followed by 250 mg every 6 hours. This medication may begiven orally, intramuscularly, or intravenously. In addition, alpha2-agonists (e.g., Apraclonidine) act by decreasing aqueous production.Their effects are additive to topically administered beta-blockers. Theyhave been approved for use in controlling an acute rise in pressurefollowing anterior chamber laser procedures, but has been reportedeffective in treating acute closed-angle glaucoma. A reasonable regimenis 1 drop every 30 minutes for 2 doses.

A second group of therapies for reducing intraocular pressure involvereducing vitreous volume. Hyperosmotic agents can be used to treat anacute attack. These agents draw water out of the globe by making theblood hyperosmolar. Oral glycerol in a dose of 1 mL/kg in a cold 50%solution (mixed with lemon juice to make it more palatable) often isused. Glycerol is converted to glucose in the liver; persons withdiabetes may need additional insulin if they become hyperglycemic afterreceiving glycerol. Oral isosorbide is a metabolically inert alcoholthat also can be used as an osmotic agent for patients with acuteangle-closure glaucoma. Usual dose is 100 g taken p.o. (220 cc of a 45%solution). This inert alcohol should not be confused with isosorbidedinitrate, a nitrate-based cardiac medication used for angina and forcongestive heart failure. Intravenous mannitol in a dose of 1.0-1.5mg/kg also is effective and is well tolerated in patients with nauseaand vomiting. These hyperosmotic agents should be used with caution inany patient with a history of congestive heart failure.

A third group of therapies involve facilitating aqueous outflow from theeye. Miotic agents pull the iris from the iridocorneal angle and mayhelp to relieve the obstruction of the trabecular meshwork by theperipheral iris. Pilocarpine 2% (blue eyes)₄% (brown eyes) can beadministered every 15 minutes for the first 1-2 hours. More frequentadministration or higher doses may precipitate a systemic cholinergiccrisis. NSAIDS are sometimes used to reduce inflammation.

Exemplary therapeutic agents for reducing intraocular pressure includeALPHAGAN® P (Allergan) (brimonidine tartrate ophthalmic solution),AZOPT® (Alcon) (brinzolamide ophthalmic suspension), BETAGAN® (Allergan)(levobunolol hydrochloride ophthalmic solution, USP), BETIMOL®(Vistakon) (timolol ophthalmic solution), BETOPTIC S® (Alcon) (betaxololHCl), BRIMONIDINE TARTRATE (Bausch & Lomb), CARTEOLOL HYDROCHLORIDE(Bausch & Lomb), COSOPT® (Merck) (dorzolamide hydrochloride-timololmaleate ophthalmic solution), LUMIGAN® (Allergan) (bimatoprostophthalmic solution), OPTIPRANOLOL® (Bausch & Lomb) (metipranololophthalmic solution), TIMOLOL GFS (Falcon) (timolol maleate ophthalmicgel forming solution), TIMOPTIC® (Merck) (timolol maleate ophthalmicsolution), TRAVATAN® (Alcon) (travoprost ophthalmic solution), TRUSOPT®(Merck) (dorzolamide hydrochloride ophthalmic solution) and XALATAN®(Pharmacia & Upjohn) (latanoprost ophthalmic solution).

In one embodiment, a sirtuin modulator can be administered inconjunction with a therapy for treating and/or preventing glaucoma. Anexample of a glaucoma drug is DARANIDE® Tablets (Merck)(Dichlorphenamide).

In one embodiment, a sirtuin modulator can be administered inconjunction with a therapy for treating and/or preventing opticneuritis. Examples of drugs for optic neuritis include DECADRON®Phosphate Injection (Merck) (Dexamethasone Sodium Phosphate),DEPO-MEDROL® (Pharmacia & Upjohn)(methylprednisolone acetate),HYDROCORTONE® Tablets (Merck) (Hydrocortisone), ORAPRED® (Biomarin)(prednisolone sodium phosphate oral solution) and PEDIAPRED® (Celltech)(prednisolone sodium phosphate, USP).

In one embodiment, a sirtuin modulator can be administered inconjunction with a therapy for treating and/or preventing CMVRetinopathy. Treatments for CMV retinopathy include CYTOVENE®(ganciclovir capsules) and VALCYTE (Roche Laboratories) (valganciclovirhydrochloride tablets).

In one embodiment, a sirtuin modulator can be administered inconjunction with a therapy for treating and/or preventing multiplesclerosis. Examples of such drugs include DANTRIUM® (Procter & GamblePharmaceuticals) (dantrolene sodium), NOVANTRONE® (Serono)(mitoxantrone), AVONEX® (Biogen Idec) (Interferon beta-1a), BETASERON®(Berlex) (Interferon beta-1b), COPAXONE® (Teva Neuroscience) (glatirameracetate injection) and REBWIF® (Pfizer) (interferon beta-1a).

In addition, macrolide and/or mycophenolic acid, which has multipleactivities, can be co-administered with a sirtuin modulator. Macrolideantibiotics include tacrolimus, cyclosporine, sirolimus, everolimus,ascomycin, erythromycin, azithromycin, clarithromycin, clindamycin,lincomycin, dirithromycin, josamycin, spiramycin, diacetyl-midecamycin,tylosin, roxithromycin, ABT-773, telithromycin, leucomycins, andlincosamide.

Mitochondrial-Associated Diseases and Disorders

In certain embodiments, the invention provides methods for treatingdiseases or disorders that would benefit from increased mitochondrialactivity. The methods involve administering to a subject in need thereofa therapeutically effective amount of a sirtuin activating compound.Increased mitochondrial activity refers to increasing activity of themitochondria while maintaining the overall numbers of mitochondria(e.g., mitochondrial mass), increasing the numbers of mitochondriathereby increasing mitochondrial activity (e.g., by stimulatingmitochondrial biogenesis), or combinations thereof. In certainembodiments, diseases and disorders that would benefit from increasedmitochondrial activity include diseases or disorders associated withmitochondrial dysfunction.

In certain embodiments, methods for treating diseases or disorders thatwould benefit from increased mitochondrial activity may compriseidentifying a subject suffering from a mitochondrial dysfunction.Methods for diagnosing a mitochondrial dysfunction may involve moleculargenetic, pathologic and/or biochemical analysis are summarized in Cohenand Gold, Cleveland Clinic Journal of Medicine, 68: 625-642 (2001). Onemethod for diagnosing a mitochondrial dysfunction is the Thor-Byrne-ierscale (see e.g., Cohen and Gold, supra; Collin S. et al., Eur Neurol.36: 260-267 (1996)). Other methods for determining mitochondrial numberand function include, for example, enzymatic assays (e.g., amitochondrial enzyme or an ATP biosynthesis factor such as an ETC enzymeor a Krebs cycle enzyme), determination or mitochondrial mass,mitochondrial volume, and/or mitochondrial number, quantification ofmitochondrial DNA, monitoring intracellular calcium homeostasis and/orcellular responses to perturbations of this homeostasis, evaluation ofresponse to an apoptogenic stimulus, determination of free radicalproduction. Such methods are known in the art and are described, forexample, in U.S. Patent Publication No. 2002/0049176 and referencescited therein.

Mitochondria are critical for the survival and proper function of almostall types of eukaryotic cells. Mitochondria in virtually any cell typecan have congenital or acquired defects that affect their function.Thus, the clinically significant signs and symptoms of mitochondrialdefects affecting respiratory chain function are heterogeneous andvariable depending on the distribution of defective mitochondria amongcells and the severity of their deficits, and upon physiological demandsupon the affected cells. Nondividing tissues with high energyrequirements, e.g. nervous tissue, skeletal muscle and cardiac muscleare particularly susceptible to mitochondrial respiratory chaindysfunction, but any organ system can be affected.

Diseases and disorders associated with mitochondrial dysfunction includediseases and disorders in which deficits in mitochondrial respiratorychain activity contribute to the development of pathophysiology of suchdiseases or disorders in a mammal. This includes 1) congenital geneticdeficiencies in activity of one or more components of the mitochondrialrespiratory chain; and 2) acquired deficiencies in the activity of oneor more components of the mitochondrial respiratory chain, wherein suchdeficiencies are caused by a) oxidative damage during aging; b) elevatedintracellular calcium; c) exposure of affected cells to nitric oxide; d)hypoxia or ischemia; e) microtubule-associated deficits in axonaltransport of mitochondria, or f) expression of mitochondrial uncouplingproteins.

Diseases or disorders that would benefit from increased mitochondrialactivity generally include for example, diseases in which free radicalmediated oxidative injury leads to tissue degeneration, diseases inwhich cells inappropriately undergo apoptosis, and diseases in whichcells fail to undergo apoptosis. Exemplary diseases or disorders thatwould benefit from increased mitochondrial activity include, forexample, AD (Alzheimer's Disease), ADPD (Alzheimer's Disease andParkinsons's Disease), AMDF (Ataxia, Myoclonus and Deafness),auto-immune disease, cancer, CIPO (Chronic Intestinal Pseudoobstructionwith myopathy and Opthalmoplegia), congenital muscular dystrophy, CPEO(Chronic Progressive External Opthalmoplegia), DEAF (Maternallyinherited DEAFness or aminoglvcoside-induced DEAFness), DEMCHO (Dementiaand Chorea), diabetes mellitus (Type I or Type II), DIDMOAD (DiabetesInsipidus, Diabetes MeHitus, Optic Atrophy, Deafness), DMDF (DiabetesMellitus and Deafness), dystonia, Exercise Intolerance, ESOC (Epilepsy,Strokes, Optic atrophy, and Cognitive decline), FBSN (Familial BilateralStriatal Necrosis), FICP (Fatal Infantile Cardiomyopathy Plus, aMELAS-associated cardiomyopathy), GER (Gastrointestinal Reflux), HD(Huntington's Disease), KSS (Kearns Sayre Syndrome), “later-onset”myopathy, LDYT (Leber's hereditary optic neuropathy and DYsTonia),Leigh's Syndrome, LHON (Leber Hereditary Optic Neuropathy), LIMM (LethalInfantile Mitochondrial Myopathy), MDM (Myopathy and Diabetes Mellitus),MELAS (Mitochondrial Encephalomyopathy, Lactic Acidosis, and Stroke-likeepisodes), MEPR (Myoclonic Epilepsy and Psychomotor Regression), MERME(MERRF/MELAS overlap disease), MERRF (Myoclonic Epilepsy and Ragged RedMuscle Fibers), MHCM (Maternally Inherited Hypertrophic CardioMyopathy),MICM (Maternally Inherited Cardiomyopathy), MILS (Maternally InheritedLeigh Syndrome), Mitochondrial Encephalocardiomyopathy, MitochondrialEncephalomyopathy, MM (Mitochondrial Myopathy), MMC (Maternal Myopathyand Cardiomyopathy), MNGIE (Myopathy and external opthalmoplegia,Neuropathy, Gastro-Intestinal, Encephalopathy), MultisystemMitochondrial Disorder (myopathy, encephalopathy, blindness, hearingloss, peripheral neuropathy), NAR^(P) (Neurogenic muscle weakness,Ataxia, and Retinitis Pigmentosa; alternate phenotype at this locus isreported as Leigh Disease), PD (Parkinson's Disease), Pearson'sSyndrome, PEM (Progressive Encephalopathy), PEO (Progressive ExternalOpthalmoplegia), PME (Progressive Myoclonus Epilepsy), PMPS (PearsonMarrow-Pancreas Syndrome), psoriasis, RTT (Rett Syndrome),schizophrenia, SIDS (Sudden Infant Death Syndrome), SNHL (SensorineuralHearing Loss), Varied Familial Presentation (clinical manifestationsrange from spastic paraparesis to multisystem progressive disorder &fatal cardiomyopathy to truncal ataxia, dysarthria, severe hearing loss,mental regression, ptosis, opthalmoparesis, distal cyclones, anddiabetes mellitus), or Wolfram syndrome.

Other diseases and disorders that would benefit from increasedmitochondrial activity include, for example, Friedreich's ataxia andother ataxias, amyotrophic lateral sclerosis (ALS) and other motorneuron diseases, macular degeneration, epilepsy, Alpers syndrome,Multiple mitochondrial DNA deletion syndrome, MtDNA depletion syndrome,Complex I deficiency, Complex II (SDH) deficiency, Complex IIIdeficiency, Cytochrome c oxidase (COX, Complex IV) deficiency, Complex Vdeficiency, Adenine Nucleotide Translocator (ANT) deficiency, Pyruvatedehydrogenase (PDH) deficiency, Ethylmalonic aciduria with lacticacidemia, 3-Methyl glutaconic aciduria with lactic acidemia, Refractoryepilepsy with declines during infection, Asperger syndrome with declinesduring infection, Autism with declines during infection, Attentiondeficit hyperactivity disorder (ADHD), Cerebral palsy with declinesduring infection, Dyslexia with declines during infection, materiallyinherited thrombocytopenia and leukemia syndrome, MARIAHS syndrome(Mitrochondrial ataxia, recurrent infections, aphasia,hypouricemia/hypomyelination, seizures, and dicarboxylic aciduria), ND6dystonia, Cyclic vomiting syndrome with declines during infection,3-Hydroxy isobutryic aciduria with lactic acidemia, Diabetes mellituswith lactic acidemia, Uridine responsive neurologic syndrome (URNS),Dilated cardiomyopathy, Splenic Lymphoma, and Renal TubularAcidosis/Diabetes/Ataxis syndrome.

In other embodiments, the invention provides methods for treating asubject suffering from mitochondrial disorders arising from, but notlimited to, post-traumatic head injury and cerebral edema, stroke(invention methods useful for preventing or preventing reperfusioninjury), Lewy body dementia, hepatorenal syndrome, acute liver failure,NASH (non-alcoholic steatohepatitis), Anti-metastasis/prodifferentiationtherapy of cancer, idiopathic congestive heart failure, atrialfibrilation (non-valvular), Wolff-Parkinson-White Syndrome, idiopathicheart block, prevention of reperfusion injury in acute myocardialinfarctions, familial migraines, irritable bowel syndrome, secondaryprevention of non-Q wave myocardial infarctions, Premenstrual syndrome,Prevention of renal failure in hepatorenal syndrome, anti-phospholipidantibody syndrome, eclampsia/pre-eclampsia, oopause infertility,ischemic heart disease/angina, and Shy-Drager and unclassifieddysautonomia syndromes.

In still another embodiment, there are provided methods for thetreatment of mitochondrial disorders associated with pharmacologicaldrug-related side effects. Types of pharmaceutical agents that areassociated with mitochondrial disorders include reverse transcriptaseinhibitors, protease inhibitors, inhibitors of DHOD, and the like.Examples of reverse transcriptase inhibitors include, for example,Azidothymidine (AZT), Stavudine (D4T), Zalcitabine (ddC), Didanosine(DDI), Fluoroiodoarauracil (FIAU), Lamivudine (3TC), Abacavir and thelike. Examples of protease inhibitors include, for example, Ritonavir,Indinavir, Saquinavir, Nelfinavir and the like. Examples of inhibitorsof dihydroorotate dehydrogenase (DHOD) include, for example,Leflunomide, Brequinar, and the like.

Reverse transcriptase inhibitors not only inhibit reverse transcriptasebut also polymerase gamma which is required for mitochondrial function.Inhibition of polymerase gamma activity (e.g., with a reversetranscriptase inhibitor) therefore leads to mitochondrial dysfunctionand/or a reduced mitochondrial mass which manifests itself in patientsas hyperlactatemia. This type of condition may benefit from an increasein the number of mitochondria and/or an improvement in mitochondrialfunction, e.g., by administration of a sirtuin activating compound.

Common symptoms of mitochondrial diseases include cardiomyopathy, muscleweakness and atrophy, developmental delays (involving motor, language,cognitive or executive function), ataxia, epilepsy, renal tubularacidosis, peripheral neuropathy, optic neuropathy, autonomic neuropathy,neurogenic bowel dysfunction, sensorineural deafness, neurogenic bladderdysfunction, dilating cardiomyopathy, migraine, hepatic failure, lacticacidemia, and diabetes mellitus.

In certain embodiments, the invention provides methods for treating adisease or disorder that would benefit from increased mitochondrialactivity that involves administering to a subject in need thereof one ormore sirtuin activating compounds in combination with anothertherapeutic agent such as, for example, an agent useful for treatingmitochondrial dysfunction (such as antioxidants, vitamins, orrespiratory chain cofactors), an agent useful for reducing a symptomassociated with a disease or disorder involving mitochondrialdysfunction (such as, an anti-seizure agent, an agent useful foralleviating neuropathic pain, an agent for treating cardiacdysfunction), a cardiovascular agent (as described further below), achemotherapeutic agent (as described further below), or ananti-neurodegeneration agent (as described further below). In anexemplary embodiment, the invention provides methods for treating adisease or disorder that would benefit from increased mitochondrialactivity that involves administering to a subject in need thereof one ormore sirtuin activating compounds in combination with one or more of thefollowing: coenzyme Q₁₀, L-carnitine, thiamine, riboflavin, niacinamide,folate, vitamin E, selenium, lipoic acid, or prednisone. Compositionscomprising such combinations are also provided herein.

In exemplary embodiments, the invention provides methods for treatingdiseases or disorders that would benefit from increased mitochondrialacitivty by administering to a subject a therapeutically effectiveamount of a sirtuin activating compound. Exemplary diseases or disordersinclude, for example, neuromuscular disorders (e.g., Friedreich'sAtaxia, muscular dystrophy, multiple sclerosis, etc.), disorders ofneuronal instability (e.g., seizure disorders, migrane, etc.),developmental delay, neurodegenerative disorders (e.g., Alzheimer'sDisease, Parkinson's Disease, amyotrophic lateral sclerosis, etc.),ischemia, renal tubular acidosis, age-related neurodegeneration andcognitive decline, chemotherapy fatigue, age-related orchemotherapy-induced menopause or irregularities of menstrual cycling orovulation, mitochondrial myopathies, mitochondrial damage (e.g., calciumaccumulation, excitotoxicity, nitric oxide exposure, hypoxia, etc.), andmitochondrial deregulation.

A gene defect underlying Friedreich's Ataxia (FA), the most commonhereditary ataxia, was recently identified and is designated “frataxin”.In FA, after a period of normal development, deficits in coordinationdevelop which progress to paralysis and death, typically between theages of 30 and 40. The tissues affected most severely are the spinalcord, peripheral nerves, myocardium, and pancreas. Patients typicallylose motor control and are confined to wheel chairs, and are commonlyafflicted with heart failure and diabetes. The genetic basis for FAinvolves GAA trinucleotide repeats in an intron region of the geneencoding frataxin. The presence of these repeats results in reducedtranscription and expression of the gene. Frataxin is involved inregulation of mitochondrial iron content. When cellular frataxin contentis subnormal, excess iron accumulates in mitochondria, promotingoxidative damage and consequent mitochondrial degeneration anddysfunction. When intermediate numbers of GAA repeats are present in thefrataxin gene intron, the severe clinical phenotype of ataxia may notdevelop. However, these intermediate-length trinucleotide extensions arefound in 25 to 30% of patients with non-insulin dependent diabetesmellitus, compared to about 5% of the nondiabetic population. In certainembodiments, sirtuin activating compounds may be used for treatingpatients with disorders related to deficiencies or defects in frataxin,including Friedreich's Ataxia, myocardial dysfunction, diabetes mellitusand complications of diabetes like peripheral neuropathy.

Muscular dystrophy refers to a family of diseases involvingdeterioration of neuromuscular structure and function, often resultingin atrophy of skeletal muscle and myocardial dysfunction. In the case ofDuchenne muscular dystrophy, mutations or deficits in a specificprotein, dystrophin, are implicated in its etiology. Mice with theirdystrophin genes inactivated display some characteristics of musculardystrophy, and have an approximately 50% deficit in mitochondrialrespiratory chain activity. A final common pathway for neuromusculardegeneration in most cases is calcium-mediated impairment ofmitochondrial function. In certain embodiments, sirtuin activatingcompounds may be used for reducing the rate of decline in muscularfunctional capacities and for improving muscular functional status inpatients with muscular dystrophy.

Multiple sclerosis (MS) is a neuromuscular disease characterized byfocal inflammatory and autoimmune degeneration of cerebral white matter.Periodic exacerbations or attacks are significantly correlated withupper respiratory tract and other infections, both bacterial and viral,indicating that mitochondrial dysfunction plays a role in MS. Depressionof neuronal mitochondrial respiratory chain activity caused by NitricOxide (produced by astrocytes and other cells involved in inflammation)is implicated as a molecular mechanism contributing to MS. In certainembodiments, sirtuin activating compounds may be used for treatment ofpatients with multiple sclerosis, both prophylactically and duringepisodes of disease exacerbation.

Epilepsy is often present in patients with mitochondrial cytopathies,involving a range of seizure severity and frequency, e.g. absence,tonic, atonic, myoclonic, and status epilepticus, occurring in isolatedepisodes or many times daily. In certain embodiments, sirtuin activatingcompounds may be used for treating patients with seizures secondary tomitochondrial dysfunction, including reducing frequency and severity ofseizure activity.

Metabolic studies on patients with recurrent migraine headaches indicatethat deficits in mitochondrial activity are commonly associated withthis disorder, manifesting as impaired-oxidative phosphorylation andexcess lactate production. Such deficits are not necessarily due togenetic defects in mitochondrial DNA. Migraineurs are hypersensitive tonitric oxide, an endogenous inhibitor of Cytochrome c Oxidase. Inaddition, patients with mitochondrial cytopathies, e.g. MELAS, oftenhave recurrent migraines. In certain embodiments, sirtuin activatingcompounds may be used for treating patients with recurrent migraineheadaches, including headaches refractory to ergot compounds orserotonin receptor antagonists.

Delays in neurological or neuropsychological development are often foundin children with mitochondrial diseases. Development and remodeling ofneural connections requires intensive biosynthetic activity,particularly involving synthesis of neuronal membranes and myelin, bothof which require pyrimidine nucleotides as cofactors. Uridinenucleotides are involved inactivation and transfer of sugars toglycolipids and glycoproteins. Cytidine nucleotides are derived fromuridine nucleotides, and are crucial for synthesis of major membranephospholipid constituents like phosphatidylcholine, which receives itscholine moiety from cytidine diphosphocholine. In the case ofmitochondrial dysfunction (due to either mitochondrial DNA defects orany of the acquired or conditional deficits like exicitoxic or nitricoxide-mediated mitochondrial dysfunction) or other conditions resultingin impaired pyrimidine synthesis, cell proliferation and axonalextension is impaired at crucial stages in development of neuronalinterconnections and circuits, resulting in delayed or arresteddevelopment of neuropsychological functions like language, motor,social, executive function, and cognitive skills. In autism for example,magnetic resonance spectroscopy measurements of cerebral phosphatecompounds indicates that there is global undersynthesis of membranes andmembrane precursors indicated by reduced levels of uridinediphospho-sugars, and cytidine nucleotide derivatives involved inmembrane synthesis. Disorders characterized by developmental delayinclude Rett's Syndrome, pervasive developmental delay (or PDD-NOS“pervasive developmental delay not otherwise specified” to distinguishit from specific subcategories like autism), autism, Asperger'sSyndrome, and Attention Deficit/Hyperactivity Disorder (ADHD), which isbecoming recognized as a delay or lag in development of neural circuitryunderlying executive functions. In certain embodiments, sirtuinactivating compounds may be useful for treating treating patients withneurodevelopmental delays (e.g., involving motor, language, executivefunction, and cognitive skills), or other delays or arrests ofneurological and neuropsychological development in the nervous systemand somatic development in non-neural tissues like muscle and endocrineglands.

The two most significant severe neurodegenerative diseases associatedwith aging, Alzheimer's Disease (AD) and Parkinson's Disease (PD), bothinvolve mitochondrial dysfunction in their pathogenesis. Complex Ideficiencies in particular are frequently found not only in thenigrostriatal neurons that degenerate in Parkinson's disease, but alsoin peripheral tissues and cells like muscle and platelets of Parkinson'sDisease patients. In Alzheimer's Disease, mitochondrial respiratorychain activity is often depressed, especially Complex IV (Cytochrome cOxidase). Moreover, mitochondrial respiratory function altogether isdepressed as a consequence of aging, further amplifying the deleterioussequelae of additional molecular lesions affecting respiratory chainfunction. Other factors in addition to primary mitochondrial dysfunctionunderlie neurodegeneration in AD, PD, and related disorders. Excitotoxicstimulation and nitric oxide are implicated in both diseases, factorswhich both exacerbate mitochondrial respiratory chain deficits and whosedeleterious actions are exaggerated on a background of respiratory chaindysfunction. Huntington's Disease also involves mitochondrialdysfunction in affected brain regions, with cooperative interactions ofexcitotoxic stimulation and mitochondrial dysfunction contributing toneuronal degeneration. In certain embodiments, sirtuin activatingcompounds may be useful for treating and attenuating progression ofage-related neurodegenerative diseases including AD and PD.

One of the major genetic defects in patients with Amyotrophic LateralSclerosis (ALS or Lou Gehrig's Disease) is mutation or deficiency inCopper-Zinc Superoxide Dismutase (SOD 1), an antioxidant enzyme.Mitochondria both produce and are primary targets for reactive oxygenspecies. Inefficient transfer of electrons to oxygen in mitochondria isthe most significant physiological source of free radicals in mammaliansystems. Deficiencies in antioxidants or antioxidant enzymes can resultin or exacerbate mitochondrial degeneration. Mice transgenic for mutatedSOD1 develop symptoms and pathology similar to those in human ALS. Thedevelopment of the disease in these animals has been shown to involveoxidative destruction of mitochondria followed by functional decline ofmotor neurons and onset of clinical symptoms. Skeletal muscle from ALSpatients has low mitochondrial Complex I activity. In certainembodiments, sirtuin activating compounds may be useful for treatingALS, for reversing or slowing the progression of clinical symptoms.

Oxygen deficiency results in both direct inhibition of mitochondrialrespiratory chain activity by depriving cells of a terminal electronacceptor for Cytochrome c reoxidation at Complex IV, and indirectly,especially in the nervous system, via secondary post-anoxicexcitotoxicity and nitric oxide formation. In conditions like cerebralanoxia, angina or sickle cell anemia crises, tissues are relativelyhypoxic. In such cases, compounds that increase mitochondrial activityprovide protection of affected tissues from deleterious effects ofhypoxia, attenuate secondary delayed cell death, and accelerate recoveryfrom hypoxic tissue stress and injury. In certain embodiments, sirtuinactivating compounds may be useful for preventing delayed cell death(apoptosis in regions like the hippocampus or cortex occurring about 2to 5 days after an episode of cerebral ischemia) after ischemic orhypoxic insult to the brain.

Acidosis due to renal dysfunction is often observed in patients withmitochondrial disease, whether the underlying respiratory chaindysfunction is congenital or induced by ischemia or cytotoxic agentslike cisplatin. Renal tubular acidosis often requires administration ofexogenous sodium bicarbonate to maintain blood and tissue pH. In certainembodiments, sirtuin activating compounds may be useful for treatingrenal tubular acidosis and other forms of renal dysfunction caused bymitochondrial respiratory chain deficits.

During normal aging, there is a progressive decline in mitochondrialrespiratory chain function. Beginning about age 40, there is anexponential rise in accumulation of mitochondrial DNA defects in humans,and a concurrent decline in nuclear-regulated elements of mitochondrialrespiratory activity. Many mitochondrial DNA lesions have a selectionadvantage during mitochondrial turnover, especially in postmitoticcells. The proposed mechanism is that mitochondria with a defectiverespiratory chain produce less oxidative damage to themselves than domitochondria with intact functional respiratory chains (mitochondrialrespiration is the primary source of free radicals in the body).Therefore, normally-functioning mitochondria accumulate oxidative damageto membrane lipids more rapidly than do defective mitochondria, and aretherefore “tagged” for degradation by lysosomes. Since mitochondriawithin cells have a half life of about 10 days, a selection advantagecan result in rapid replacement of functional mitochondria with thosewith diminished respiratory activity, especially in slowly dividingcells. The net result is that once a mutation in a gene for amitochondrial protein that reduces oxidative damage to mitochondriaoccurs, such defective mitochondria will rapidly populate the cell,diminishing or eliminating its respiratory capabilities. Theaccumulation of such cells results in aging or degenerative disease atthe organismal level. This is consistent with the progressive mosaicappearance of cells with defective electron transport activity inmuscle, with cells almost devoid of Cytochrome c Oxidase (COX) activityinterspersed randomly amidst cells with normal activity, and a higherincidence of COX-negative cells in biopsies from older subjects. Theorganism, during aging, or in a variety of mitochondrial diseases, isthus faced with a situation in which irreplaceable postmitotic cells(e.g. neurons, skeletal and cardiac muscle) must be preserved and theirfunction maintained to a significant degree, in the face of aninexorable progressive decline in mitochondrial respiratory chainfunction. Neurons with dysfunctional mitochondria become progressivelymore sensitive to insults like excitotoxic injury. Mitochondrial failurecontributes to most degenerative diseases (especially neurodegeneration)that accompany aging. Congenital mitochondrial diseases often involveearly-onset neurodegeneration similar in fundamental mechanism todisorders that occur during aging of people born with normalmitochondria. In certain embodiments, sirtuin activating compounds maybe useful for treating or attenuating cognitive decline and otherdegenerative consequences of aging.

Mitochondrial DNA damage is more extensive and persists longer thannuclear DNA damage in cells subjected to oxidative stress or cancerchemotherapy agents like cisplatin due to both greater vulnerability andless efficient repair of mitochondrial DNA. Although mitochondrial DNAmay be more sensitive to damage than nuclear DNA, it is relativelyresistant, in some situations, to mutagenesis by chemical carcinogens.This is because mitochondria respond to some types of mitochondrial DNAdamage by destroying their defective genomes rather than attempting torepair them. This results in global mitochondrial dysfunction for aperiod after cytotoxic chemotherapy. Clinical use of chemotherapy agentslike cisplatin, mitomycin, and cytoxan is often accompanied bydebilitating “chemotherapy fatigue”, prolonged periods of weakness andexercise intolerance which may persist even after recovery fromhematologic and gastrointestinal toxicities of such agents. In certainembodiments, sirtuin activating compounds may be useful for treatmentand prevention of side effects of cancer chemotherapy related tomitochondrial dysfunction.

A crucial function of the ovary is to maintain integrity of themitochondrial genome in oocytes, since mitochondria passed onto a fetusare all derived from those present in oocytes at the time of conception.Deletions in mitochondrial DNA become detectable around the age ofmenopause, and are also associated with abnormal menstrual cycles. Sincecells cannot directly detect and respond to defects in mitochondrialDNA, but can only detect secondary effects that affect the cytoplasm,like impaired respiration, redox status, or deficits in pyrimidinesynthesis, such products of mitochondrial function participate as asignal for oocyte selection and follicular atresia, ultimatelytriggering menopause when maintenance of mitochondrial genomic fidelityand functional activity can no longer be guaranteed. This is analogousto apoptosis in cells with DNA damage, which undergo an active processof cellular suicide when genomic fidelity can no longer be achieved byrepair processes. Women with mitochondrial cytopathies affecting thegonads often undergo premature menopause or display primary cyclingabnormalities. Cytotoxic cancer chemotherapy often induces prematuremenopause, with a consequent increased risk of osteoporosis.Chemotherapy-induced amenorrhea is generally due to primary ovarianfailure. The incidence of chemotherapy-induced amenorrhea increases as afunction of age in premenopausal women receiving chemotherapy, pointingtoward mitochondrial involvement. Inhibitors of mitochondrialrespiration or protein synthesis inhibit hormone-induced ovulation, andfurthermore inhibit production of ovarian steroid hormones in responseto pituitary gonadotropins. Women with Down's syndrome typically undergomenopause prematurely, and also are subject to early onset ofAlzheimer-like dementia. Low activity of cytochrome oxidase isconsistently found in tissues of Down's patients and in late-onsetAlzheimer's Disease. Appropriate support of mitochondrial function orcompensation for mitochondrial dysfunction therefore is useful forprotecting against age-related or chemotherapy-induced menopause orirregularities of menstrual cycling or ovulation. In certainembodiments, sirtuin activating compounds may be useful for treating andpreventing amenorrhea, irregular ovulation, menopause, or secondaryconsequences of menopause.

In certain embodiments, sirtuin modulating compounds may be useful fortreatment mitochondrial myopathies. Mitochondrial myopathies range frommild, slowly progressive weakness of the extraocular muscles to severe,fatal infantile myopathies and multisystem encephalomyopathies. Somesyndromes have been defined, with some overlap between them. Establishedsyndromes affecting muscle include progressive external opthalmoplegia,the Kearns-Sayre syndrome (with opthalmoplegia, pigmentary retinopathy,cardiac conduction defects, cerebellar ataxia, and sensorineuraldeafness), the MELAS syndrome (mitochondrial encephalomyopathy, lacticacidosis, and stroke-like episodes), the MERFF syndrome (myoclonicepilepsy and ragged red fibers), limb-girdle distribution weakness, andinfantile myopathy (benign or severe and fatal). Muscle biopsy specimensstained with modified Gomori's trichrome stain show ragged red fibersdue to excessive accumulation of mitochondria. Biochemical defects insubstrate transport and utilization, the Krebs cycle, oxidativephosphorylation, or the respiratory chain are detectable. Numerousmitochondrial DNA point mutations and deletions have been described,transmitted in a maternal, nonmendelian inheritance pattern. Mutationsin nuclear-encoded mitochondrial enzymes occur.

In certain embodiments, sirtuin activating compounds may be useful fortreating patients suffering from toxic damage to mitochondria, such as,toxic damage due to calcium accumulation, excitotoxicity, nitric oxideexposure, drug induced toxic damage, or hypoxia.

A fundamental mechanism of cell injury, especially in excitable tissues,involves excessive calcium entry into cells, as a result of eitherleakage through the plasma membrane or defects in intracellular calciumhandling mechanisms. Mitochondria are major sites of calciumsequestration, and preferentially utilize energy from the respiratorychain for taking up calcium rather than for ATP synthesis, which resultsin a downward spiral of mitochondrial failure, since calcium uptake intomitochondria results in diminished capabilities for energy transduction.

Excessive stimulation of neurons with excitatory amino acids is a commonmechanism of cell death or injury in the central nervous system.Activation of glutamate receptors, especially of the subtype designatedNMDA receptors, results in mitochondrial dysfunction, in part throughelevation of intracellular calcium during excitotoxic stimulation.Conversely, deficits in mitochondrial respiration and oxidativephosphorylation sensitizes cells to excitotoxic stimuli, resulting incell death or injury during exposure to levels of excitotoxicneurotransmitters or toxins that would be innocuous to normal cells.

Nitric oxide (about 1 micromolar) inhibits cytochrome oxidase (ComplexIV) and thereby inhibits mitochondrial respiration; moreover, prolongedexposure to nitric oxide (NO) irreversibly reduces Complex I activity.Physiological or pathophysiological concentrations of NO thereby inhibitpyrimidine biosynthesis. Nitric oxide is implicated in a variety ofneurodegenerative disorders including inflammatory and autoimmunediseases of the central nervous system, and is involved in mediation ofexcitotoxic and post-hypoxic damage to neurons.

Oxygen is the terminal electron acceptor in the respiratory chain.Oxygen deficiency impairs electron transport chain activity, resultingin diminished pyrimidine synthesis as well as diminished ATP synthesisvia oxidative phosphorylation. Human cells proliferate and retainviability under virtually anaerobic conditions if provided with uridineand pyruvate (or a similarly effective agent for oxidizing NADH tooptimize glycolytic ATP production).

In certain embodiments, sirtuin activating compounds may be useful fortreating diseases or disorders associated with mitochondrialderegulation.

Transcription of mitochondrial DNA encoding respiratory chain componentsrequires nuclear factors. In neuronal axons, mitochondria must shuttleback and forth to the nucleus in order to maintain respiratory chainactivity. If axonal transport is impaired by hypoxia or by drugs liketaxol which affect microtubule stability, mitochondria distant from thenucleus undergo loss of cytochrome oxidase activity. Accordingly,treatment with a sirtuin activating compound may be useful for promotingnuclear-mitochondrial interactions.

Mitochondria are the primary source of free radicals and reactive oxygenspecies, due to spillover from the mitochondrial respiratory chain,especially when defects in one or more respiratory chain componentsimpairs orderly transfer of electrons from metabolic intermediates tomolecular oxygen. To reduce oxidative damage, cells can compensate byexpressing mitochondrial uncoupling proteins (UCP), of which severalhave been identified. UCP-2 is transcribed in response to oxidativedamage, inflammatory cytokines, or excess lipid loads, e.g. fatty liverand steatohepatitis. UCPs reduce spillover of reactive oxygen speciesfrom mitochondria by discharging proton gradients across themitochondrial inner membrane, in effect wasting energy produced bymetabolism and rendering cells vulnerable to energy stress as atrade-off for reduced oxidative injury.

Muscle Performance

In other embodiments, the invention provides methods for enhancingmuscle performance by administering a therapeutically effective amountof a sirtuin activating compound. For example, sirtuin activatingcompounds may be useful for improving physical endurance (e.g., abilityto perform a physical task such as exercise, physical labor, sportsactivities, etc.), inhibiting or retarding physical fatigues, enhancingblood oxygen levels, enhancing energy in healthy individuals, enhanceworking capacity and endurance, reducing muscle fatigue, reducingstress, enhancing cardiac and cardiovascular function, improving sexualability, increasing muscle ATP levels, and/or reducing lactic acid inblood. In certain embodiments, the methods involve administering anamount of a sirtuin activating compound that increase mitochondrialactivity, increase mitochondrial biogenesis, and/or increasemitochondrial mass.

Sports performance refers to the ability of the athlete's muscles toperform when participating in sports activities. Enhanced sportsperformance, strength, speed and endurance are measured by an increasein muscular contraction strength, increase in amplitude of musclecontraction, shortening of muscle reaction time between stimulation andcontraction. Athlete refers to an individual who participates in sportsat any level and who seeks to achieve an improved level of strength,speed and endurance in their performance, such as, for example, bodybuilders, bicyclists, long distance runners, short distance runners,etc. An athlete may be hard training, that is, performs sportsactivities intensely more than three days a week or for competition. Anathlete may also be a fitness enthusiast who seeks to improve generalhealth and well-being, improve energy levels, who works out for about1-2 hours about 3 times a week. Enhanced sports performance inmanifested by the ability to overcome muscle fatigue, ability tomaintain activity for longer periods of time, and have a more effectiveworkout.

In the arena of athlete muscle performance, it is desirable to createconditions that permit competition or training at higher levels ofresistance for a prolonged period of time. However, acute and intenseanaerobic use of skeletal muscles often results in impaired athleticperformance, with losses in force and work output, and increased onsetof muscle fatigue, soreness, and dysfunction. It is now recognized thateven a single exhaustive exercise session, or for that matter any acutetrauma to the body such as muscle injury, resistance or exhaustivemuscle exercise, or elective surgery, is characterized by perturbedmetabolism that affects muscle performance in both short and long termphases. Both muscle metabolic/enzymatic activity and gene expression areaffected. For example, disruption of skeletal muscle nitrogen metabolismas well as depletion of sources of metabolic energy occur duringextensive muscle activity. Amino acids, including branched-chain aminoacids, are released from muscles followed by their deamination toelevate serum ammonia and local oxidation as muscle fuel sources, whichaugments metabolic acidosis. In addition, there is a decline incatalytic efficiency of muscle contraction events, as well as analteration of enzymatic activities of nitrogen and energy metabolism.Further, protein catabolism is initiated where rate of protein synthesisis decreased coupled with an increase in the degradation ofnon-contractible protein. These metabolic processes are also accompaniedby free radical generation which further damages muscle cells.

Recovery from fatigue during acute and extended exercise requiresreversal of metabolic and non-metabolic fatiguing factors. Known factorsthat participate in human muscle fatigue, such as lactate, ammonia,hydrogen ion, etc., provide an incomplete and unsatisfactory explanationof the fatigue/recovery process, and it is likely that additionalunknown agents participate (Baker et al., J. Appl. Physiol.74:2294-2300, 1993; Bazzarre et al., J. Am. Coll. Nutr. 11:505-511,1992; Dohm et al., Fed. Proc. 44:348-352, 1985; Edwards In: Biochemistryof Exercise, Proceedings of the Fifth International Symposium on theBiochemistry of Exercise (Kutrgen, Vogel, Poormans, eds.), 1983;MacDougall et al., Acta Physiol. Scand. 146:403-404, 1992; Walser etal., Kidney Int. 32:123-128, 1987). Several studies have also analyzedthe effects of nutritional supplements and herbal supplements inenhancing muscle performance.

Aside from muscle performance during endurance exercise, free radicalsand oxidative stress parameters are affected in pathophysiologicalstates. A substantial body of data now suggests that oxidative stresscontributes to muscle wasting or atrophy in pathophysiological states(reviewed in Clarkson, P. M. Antioxidants and physical performance.Crit. Rev. Food Sci. Nutr. 35: 3141; 1995; Powers, S. K.; Lennon, S. L.Analysis of cellular responses to free radicals: Focus on exercise andskeletal muscle. Proc. Nutr. Soc. 58: 1025-1033; 1999). For example,with respect to muscular disorders where both muscle endurance andfunction are compensated, the role of nitric oxide (NO), has beenimplicated. In muscular dystrophies, especially those due to defects inproteins that make up the dystrophin-glycoprotein complex (DGC), theenzyme that synthesizes NO, nitric oxide synthase (NOS), has beenassociated. Recent studies of dystrophies related to DGC defects suggestthat one mechanism of cellular injury is functional ischemia related toalterations in cellular NOS and disruption of a normal protective actionof NO. This protective action is the prevention of local ischemia duringcontraction-induced increases in sympathetic vasoconstriction. Rando(Microsc Res Tech 55(4):223-35, 2001), has shown that oxidative injuryprecedes pathologic changes and that muscle cells with defects in theDGC have an increased susceptibility to oxidant challenges. Excessivelipid peroxidation due to free radicals has also been shown to be afactor in myopathic diseases such as McArdle's disease (Russo et al.,Med. Hypotheses. 39(2):147-51, 1992). Furthermore, mitochondrialdysfunction is a well-known correlate of age-related muscle wasting(sarcopenia) and free radical damage has been suggested, though poorlyinvestigated, as a contributing factor (reviewed in Navarro, A.;Lopez-Cepero, J. M.; Sanchez del Pino, M. L. Front. Biosci. 6: D2644;2001). Other indications include acute sarcopenia, for example muscleatrophy and/or cachexia associated with burns, bed rest, limbimmobilization, or major thoracic, abdominal, and/or orthopedic surgery.It is contemplated that the methods of the present invention will alsobe effective in the treatment of muscle related pathological conditions.

In certain embodiments, the invention provides novel dietarycompositions comprising sirtuin modulators, a method for theirpreparation, and a method of using the compositions for improvement ofsports performance. Accordingly, provided are therapeutic compositions,foods and beverages that have actions of improving physical enduranceand/or inhibiting physical fatigues for those people involved inbroadly-defined exercises including sports requiring endurance andlabors requiring repeated muscle exertions. Such dietary compositionsmay additional comprise electrolytes, caffeine, vitamins, carbohydrates,etc.

Other Uses

Sirtuin-modulating compounds that increase the level and/or activity ofa sirtuin protein may be used for treating or preventing viralinfections (such as infections by influenza, herpes or papilloma virus)or as antifungal agents. In certain embodiments, sirtuin-modulatingcompounds that increase the level and/or activity of a sirtuin proteinmay be administered as part of a combination drug therapy with anothertherapeutic agent for the treatment of viral diseases, including, forexample, acyclovir, ganciclovir and zidovudine. In another embodiment,sirtuin-modulating compounds that increase the level and/or activity ofa sirtuin protein may be administered as part of a combination drugtherapy with another anti-fungal agent including, for example, topicalanti-fungals such as ciclopirox, clotrimazole, econazole, miconazole,nystatin, oxiconazole, terconazole, and tolnaftate, or systemicanti-fungal such as fluconazole (Diflucan), itraconazole (Sporanox),ketoconazole (Nizoral), and miconazole (Monistat I.V.).

Subjects that may be treated as described herein include eukaryotes,such as mammals, e.g., humans, ovines, bovines, equines, porcines,canines, felines, non-human primate, mice, and rats. Cells that may betreated include eukaryotic cells, e.g., from a subject described above,or plant cells, yeast cells and prokaryotic cells, e.g., bacterialcells. For example, modulating compounds may be administered to farmanimals to improve their ability to withstand farming conditions longer.

Sirtuin-modulating compounds that increase the level and/or activity ofa sirtuin protein may also be used to increase lifespan, stressresistance, and resistance to apoptosis in plants. In one embodiment, acompound is applied to plants, e.g., on a periodic basis, or to fungi.In another embodiment, plants are genetically modified to produce acompound. In another embodiment, plants and fruits are treated with acompound prior to picking and shipping to increase resistance to damageduring shipping. Plant seeds may also be contacted with compoundsdescribed herein, e.g., to preserve them.

In other embodiments, sirtuin-modulating compounds that increase thelevel and/or activity of a sirtuin protein may be used for modulatinglifespan in yeast cells. Situations in which it may be desirable toextend the lifespan of yeast cells include any process in which yeast isused, e.g., the making of beer, yogurt, and bakery items, e.g., bread.Use of yeast having an extended lifespan can result in using less yeastor in having the yeast be active for longer periods of time. Yeast orother mammalian cells used for recombinantly producing proteins may alsobe treated as described herein.

Sirtuin-modulating compounds that increase the level and/or activity ofa sirtuin protein may also be used to increase lifespan, stressresistance and resistance to apoptosis in insects. In this embodiment,compounds would be applied to useful insects, e.g., bees and otherinsects that are involved in pollination of plants. In a specificembodiment, a compound would be applied to bees involved in theproduction of honey. Generally, the methods described herein may beapplied to any organism, e.g., eukaryote, that may have commercialimportance. For example, they can be applied to fish (aquaculture) andbirds (e.g., chicken and fowl).

Higher doses of sirtuin-modulating compounds that increase the leveland/or activity of a sirtuin protein may also be used as a pesticide byinterfering with the regulation of silenced genes and the regulation ofapoptosis during development. In this embodiment, a compound may beapplied to plants using a method known in the art that ensures thecompound is bio-available to insect larvae, and not to plants.

At least in view of the link between reproduction and longevity (Longoand Finch, Science, 2002), sirtuin-modulating compounds that increasethe level and/or activity of a sirtuin protein can be applied to affectthe reproduction of organisms such as insects, animals andmicroorganisms.

4. Assays

Yet other methods contemplated herein include screening methods foridentifying compounds or agents that modulate sirtuins. An agent may bea nucleic acid, such as an aptamer. Assays may be conducted in a cellbased or cell free format. For example, an assay may comprise incubating(or contacting) a sirtuin with a test agent under conditions in which asirtuin can be modulated by an agent known to modulate the sirtuin, andmonitoring or determining the level of modulation of the sirtuin in thepresence of the test agent relative to the absence of the test agent.The level of modulation of a sirtuin can be determined by determiningits ability to deacetylate a substrate. Exemplary substrates areacetylated peptides which can be obtained from BIOMOL (Plymouth Meeting,Pa.). Preferred substrates include peptides of p53, such as thosecomprising an acetylated K382. A particularly preferred substrate is theFluor de Lys-SIRT1 (BIOMOL), i.e., the acetylated peptideArg-His-Lys-Lys. Other substrates are peptides from human histones H3and H4 or an acetylated amino acid. Substrates may be fluorogenic. Thesirtuin may be SIRT1, Sir2, SIRT3, or a portion thereof. For example,recombinant SIRT1 can be obtained from BIOMOL. The reaction may beconducted for about 30 minutes and stopped, e.g., with nicotinamide. TheHDAC fluorescent activity assay/drug discovery kit (AK-500, BIOMOLResearch Laboratories) may be used to determine the level ofacetylation. Similar assays are described in Bitterman et al. (2002) J.Biol. Chem. 277:45099. The level of modulation of the sirtuin in anassay may be compared to the level of modulation of the sirtuin in thepresence of one or more (separately or simultaneously) compoundsdescribed herein, which may serve as positive or negative controls.Sirtuins for use in the assays may be full length sirtuin proteins orportions thereof. Since it has been shown herein that activatingcompounds appear to interact with the N-terminus of SIRT1, proteins foruse in the assays include N-terminal portions of sirtuins, e.g., aboutamino acids 1-176 or 1-255 of SIRT1; about amino acids 1-174 or 1-252 ofSir2.

In one embodiment, a screening assay comprises (i) contacting a sirtuinwith a test agent and an acetylated substrate under conditionsappropriate for the sirtuin to deacetylate the substrate in the absenceof the test agent; and (ii) determining the level of acetylation of thesubstrate, wherein a lower level of acetylation of the substrate in thepresence of the test agent relative to the absence of the test agentindicates that the test agent stimulates deacetylation by the sirtuin,whereas a higher level of acetylation of the substrate in the presenceof the test agent relative to the absence of the test agent indicatesthat the test agent inhibits deacetylation by the sirtuin.

Methods for identifying an agent that modulates, e.g., stimulates orinhibits, sirtuins in vivo may comprise (i) contacting a cell with atest agent and a substrate that is capable of entering a cell in thepresence of an inhibitor of class I and class II HDACs under conditionsappropriate for the sirtuin to deacetylate the substrate in the absenceof the test agent; and (ii) determining the level of acetylation of thesubstrate, wherein a lower level of acetylation of the substrate in thepresence of the test agent relative to the absence of the test agentindicates that the test agent stimulates deacetylation by the sirtuin,whereas a higher level of acetylation of the substrate in the presenceof the test agent relative to the absence of the test agent indicatesthat the test agent inhibits deacetylation by the sirtuin. A preferredsubstrate is an acetylated peptide, which is also preferablyfluorogenic, as further described herein. The method may furthercomprise lysing the cells to determine the level of acetylation of thesubstrate. Substrates may be added to cells at a concentration rangingfrom about 1 μM to about 10 mM, preferably from about 10 μM to 1 mM,even more preferably from about 100 μM to 1 mM, such as about 200 μM. Apreferred substrate is an acetylated lysine, e.g., ε-acetyl lysine(Fluor de Lys, FdL) or Fluor de Lys-SIRT1. A preferred inhibitor ofclass I and class II HDACs is trichostatin A (TSA), which may be used atconcentrations ranging from about 0.01 to 100 μM, preferably from about0.1 to 10 μM, such as 1 μM. Incubation of cells with the test compoundand the substrate may be conducted for about 10 minutes to 5 hours,preferably for about 1-3 hours. Since TSA inhibits all class I and classII HDACs, and that certain substrates, e.g., Fluor de Lys, is a poorsubstrate for SIRT2 and even less a substrate for SIRT3-7, such an assaymay be used to identify modulators of SIRT1 in vivo.

5. Pharmaceutical Compositions

The sirtuin-modulating compounds described herein may be formulated in aconventional manner using one or more physiologically acceptablecarriers or excipients. For example, sirtuin-modulating compounds andtheir physiologically acceptable salts and solvates may be formulatedfor administration by, for example, injection (e.g. SubQ, IM, IP),inhalation or insufflation (either through the mouth or the nose) ororal, buccal, sublingual, transdermal, nasal, parenteral or rectaladministration. In one embodiment, a sirtuin-modulating compound may beadministered locally, at the site where the target cells are present,i.e., in a specific tissue, organ, or fluid (e.g., blood, cerebrospinalfluid, etc.).

Sirtuin-modulating compounds can be formulated for a variety of modes ofadministration, including systemic and topical or localizedadministration. Techniques and formulations generally may be found inRemington's Pharmaceutical Sciences, Meade Publishing Co., Easton, Pa.For parenteral administration, injection is preferred, includingintramuscular, intravenous, intraperitoneal, and subcutaneous. Forinjection, the compounds can be formulated in liquid solutions,preferably in physiologically compatible buffers such as Hank's solutionor Ringer's solution. In addition, the compounds may be formulated insolid form and redissolved or suspended immediately prior to use.Lyophilized forms are also included.

For oral administration, the pharmaceutical compositions may take theform of, for example, tablets, lozanges, or capsules prepared byconventional means with pharmaceutically acceptable excipients such asbinding agents (e.g., pregelatinised maize starch, polyvinylpyrrolidoneor hydroxypropyl methylcellulose); fillers (e.g., lactose,microcrystalline cellulose or calcium hydrogen phosphate); lubricants(e.g., magnesium stearate, talc or silica); disintegrants (e.g., potatostarch or sodium starch glycolate); or wetting agents (e.g., sodiumlauryl sulphate). The tablets may be coated by methods well known in theart. Liquid preparations for oral administration may take the form of,for example, solutions, syrups or suspensions, or they may be presentedas a dry product for constitution with water or other suitable vehiclebefore use. Such liquid preparations may be prepared by conventionalmeans with pharmaceutically acceptable additives such as suspendingagents (e.g., sorbitol syrup, cellulose derivatives or hydrogenatededible fats); emulsifying agents (e.g., lecithin or acacia); non-aqueousvehicles (e.g., ationd oil, oily esters, ethyl alcohol or fractionatedvegetable oils); and preservatives (e.g., methyl orpropyl-p-hydroxybenzoates or sorbic acid). The preparations may alsocontain buffer salts, flavoring, coloring and sweetening agents asappropriate. Preparations for oral administration may be suitablyformulated to give controlled release of the active compound.

For administration by inhalation (e.g., pulmonary delivery),sirtuin-modulating compounds may be conveniently delivered in the formof an aerosol spray presentation from pressurized packs or a nebuliser,with the use of a suitable propellant, e.g., dichlorodifluoromethane,trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide orother suitable gas. In the case of a pressurized aerosol the dosage unitmay be determined by providing a valve to deliver a metered amount.Capsules and cartridges of e.g., gelatin, for use in an inhaler orinsufflator may be formulated containing a powder mix of the compoundand a suitable powder base such as lactose or starch.

Sirtuin-modulating compounds may be formulated for parenteraladministration by injection, e.g., by bolus injection or continuousinfusion. Formulations for injection may be presented in unit dosageform, e.g., in ampoules or in multi-dose containers, with an addedpreservative. The compositions may take such forms as suspensions,solutions or emulsions in oily or aqueous vehicles, and may containformulatory agents such as suspending, stabilizing and/or dispersingagents. Alternatively, the active ingredient may be in powder form forconstitution with a suitable vehicle, e.g., sterile pyrogen-free water,before use.

Sirtuin-modulating compounds may also be formulated in rectalcompositions such as suppositories or retention enemas, e.g., containingconventional suppository bases such as cocoa butter or other glycerides.

In addition to the formulations described previously, sirtuin-modulatingcompounds may also be formulated as a depot preparation. Such longacting formulations may be administered by implantation (for examplesubcutaneously or intramuscularly) or by intramuscular injection. Thus,for example, sirtuin-modulating compounds may be formulated withsuitable polymeric or hydrophobic materials (for example as an emulsionin an acceptable oil) or ion exchange resins, or as sparingly solublederivatives, for example, as a sparingly soluble salt. Controlledrelease formula also includes patches.

In certain embodiments, the compounds described herein can be formulatedfor delivery to the central nervous system (CNS) (reviewed in Begley,Pharmacology & Therapeutics 104: 2945 (2004)). Conventional approachesfor drug delivery to the CNS include: neurosurgical strategies (e.g.,intracerebral injection or intracerebroventricular infusion); molecularmanipulation of the agent (e.g., production of a chimeric fusion proteinthat comprises a transport peptide that has an affinity for anendothelial cell surface molecule in combination with an agent that isitself incapable of crossing the BBB) in an attempt to exploit one ofthe endogenous transport pathways of the BBB; pharmacological strategiesdesigned to increase the lipid solubility of an agent (e.g., conjugationof water-soluble agents to lipid or cholesterol carriers); and thetransitory disruption of the integrity of the BBB by hyperosmoticdisruption (resulting from the infusion of a mannitol solution into thecarotid artery or the use of a biologically active agent such as anangiotensin peptide).

One possibility to achieve sustained release kinetics is embedding orencapsulating the active compound into nanoparticles. Nanoparticles canbe administrated as powder, as a powder mixture with added excipients oras suspensions. Colloidal suspensions of nanoparticles can easily beadministrated through a cannula with small diameter.

Nanoparticles are particles with a diameter from about 5 nm to up toabout 1000 nm. The term “nanoparticles” as it is used hereinafter refersto particles formed by a polymeric matrix in which the active compoundis dispersed, also known as “nanospheres”, and also refers tonanoparticles which are composed of a core containing the activecompound which is surrounded by a polymeric membrane, also known as“nanocapsules”. In certain embodiments, nanoparticles are preferredhaving a diameter from about 50 nm to about 500 nm, in particular fromabout 100 nm to about 200 nm.

Nanoparticles can be prepared by in situ polymerization of dispersedmonomers or by using preformed polymers. Since polymers prepared in situare often not biodegradable and/or contain toxicological seriousbyproducts, nanoparticles from preformed polymers are preferred.Nanoparticles from preformed polymers can be prepared by differenttechniques, e.g., by emulsion evaporation, solvent displacement,salting-out, mechanical grinding, microprecipitation, and byemulsification diffusion.

With the methods described above, nanoparticles can be formed withvarious types of polymers. For use in the method of the presentinvention, nanoparticles made from biocompatible polymers are preferred.The term “biocompatible” refers to material that after introduction intoa biological environment has no serious effects to the biologicalenvironment. From biocompatible polymers those polymers are especiallypreferred which are also biodegradable. The term “biodegradable” refersto material that after introduction into a biological environment isenzymatically or chemically degraded into smaller molecules, which canbe eliminated subsequently. Examples are polyesters fromhydroxycarboxylic acids such as poly(lactic acid) (PLA), poly(glycolicacid) (PGA), polycaprolactone (PCL), copolymers of lactic acid andglycolic acid (PLGA), copolymers of lactic acid and caprolactone,polyepsilon caprolactone, polyhyroxy butyric acid and poly(ortho)esters,polyurethanes, polyanhydrides, polyacetals, polydihydropyrans,polycyanoacrylates, natural polymers such as alginate and otherpolysaccharides including dextran and cellulose, collagen and albumin.

Suitable surface modifiers can preferably be selected from known organicand inorganic pharmaceutical excipients. Such excipients include variouspolymers, low molecular weight oligomers, natural products andsurfactants. Preferred surface modifiers include nonionic and ionicsurfactants. Representative examples of surface modifiers includegelatin, casein, lecithin (phosphatides), gum acacia, cholesterol,tragacanth, stearic acid, benzalkonium chloride, calcium stearate,glycerol monostearate, cetostearyl alcohol, cetomacrogol emulsifyingwax, sorbitan esters, polyoxyethylene alkyl ethers, e.g., macrogolethers such as cetomacrogol 1000, polyoxyethylene castor oilderivatives, polyoxyethylene sorbitan fatty acid esters, e.g., thecommercially available Tweens™, polyethylene glycols, polyoxyethylenestearates, colloidal silicon dioxide, phosphates, sodium dodecylsulfate,carboxymethylcellulose calcium, carboxymethylcellulose sodium,methylcellulose, hydroxyethylcellulose, hydroxy propylcellulose,hydroxypropylmethylcellulose phthalate, noncrystalline cellulose,magnesium aluminum silicate, triethanolamine, polyvinyl alcohol, andpolyvinylpyrrolidone (PVP). Most of these surface modifiers are knownpharmaceutical excipients and are described in detail in the Handbook ofPharmaceutical Excipients, published jointly by the AmericanPharmaceutical Association and The Pharmaceutical Society of GreatBritain, the Pharmaceutical Press, 1986.

Further description on preparing nanoparticles can be found, forexample, in U.S. Pat. No. 6,264,922, the contents of which areincorporated herein by reference.

Liposomes are a further drug delivery system which is easily injectable.Accordingly, in the method of invention the active compounds can also beadministered in the form of a liposome delivery system. Liposomes arewell-known by a person skilled in the art. Liposomes can be formed froma variety of phospholipids, such as cholesterol, stearylamine ofphosphatidylcholines. Liposomes being usable for the method of inventionencompass all types of liposomes including, but not limited to, smallunilamellar vesicles, large unilamellar vesicles and multilamellarvesicles.

Liposomes are used for a variety of therapeutic purposes, and inparticular, for carrying therapeutic agents to target cells.Advantageously, liposome-drug formulations offer the potential ofimproved drug-delivery properties, which include, for example,controlled drug release. An extended circulation time is often neededfor liposomes to reach a target region, cell or site. In particular,this is necessary where the target region, cell or site is not locatednear the site of administration. For example, when liposomes areadministered systemically, it is desirable to coat the liposomes with ahydrophilic agent, for example, a coating of hydrophilic polymer chainssuch as polyethylene glycol (PEG) to extend the blood circulationlifetime of the liposomes. Such surface-modified liposomes are commonlyreferred to as “long circulating” or “sterically stabilized” liposomes.

One surface modification to a liposome is the attachment of PEG chains,typically having a molecular weight from about 1000 daltons (Da) toabout 5000 Da, and to about 5 mole percent (%) of the lipids making upthe liposomes (see, for example, Stealth Liposomes, CRC Press, Lasic, D.and Martin, F., eds., Boca Raton, Fla., (1995)), and the citedreferences therein. The pharmacokinetics exhibited by such liposomes arecharacterized by a dose-independent reduction in uptake of liposomes bythe liver and spleen via the mononuclear phagocyte system (MPS), andsignificantly prolonged blood circulation time, as compared tonon-surface-modified liposomes, which tend to be rapidly removed fromthe blood and accumulated in the liver and spleen.

In certain embodiments, the complex is shielded to increase thecirculatory half-life of the complex or shielded to increase theresistance of nucleic acid to degradation, for example degradation bynucleases.

As used herein, the term “shielding”, and its cognates such as“shielded”, refers to the ability of “shielding moieties” to reduce thenon-specific interaction of the complexes described herein with serumcomplement or with other species present in serum in vitro or in vivo.Shielding moieties may decrease the complex interaction with or bindingto these species through one or more mechanisms, including, for example,non-specific steric or non-specific electronic interactions. Examples ofsuch interactions include non-specific electrostatic interactions,charge interactions, Van der Waals interactions, steric-hindrance andthe like. For a moiety to act as a shielding moiety, the mechanism ormechanisms by which it may reduce interaction with, association with orbinding to the serum complement or other species does not have to beidentified. One can determine whether a moiety can act as a shieldingmoiety by determining whether or to what extent a complex binds serumspecies.

It should be noted that “shielding moieties” can be multifunctional. Forexample, a shielding moiety may also function as, for example, atargeting factor. A shielding moiety may also be referred to asmultifunctional with respect to the mechanism(s) by which it shields thecomplex. While not wishing to be limited by proposed mechanism ortheory, examples of such a multifunctional shielding moiety are pHsensitive endosomal membrane-disruptive synthetic polymers, such as PPAAor PEAA. Certain poly(alkylacrylic acids) have been shown to disruptendosomal membranes while leaving the-outer cell surface membrane intact(Stayton et al. (2000) J. Controll. Release 65:203-220; Murthy et al.(1999) J. Controll. Release 61:137-143; WO 99/34831), thereby increasingcellular bioavailability and functioning as a targeting factor. However,PPAA reduces binding of serum complement to complexes in which it isincorporated, thus functioning as a shielding moiety.

Another way to produce a formulation, particularly a solution, of asirtuin modulator such as resveratrol or a derivative thereof, isthrough the use of cyclodextrin. By cyclodextrin is meant α-, β-, orγ-cyclodextrin. Cyclodextrins are described in detail in Pitha et al.,U.S. Pat. No. 4,727,064, which is incorporated herein by reference.Cyclodextrins are cyclic oligomers of glucose; these compounds forminclusion complexes with any drug whose molecule can fit into thelipophile-seeking cavities of the cyclodextrin molecule.

The cyclodextrin of the compositions according to the invention may beα-, β-, or γ-cyclodextrin. α-cyclodextrin contains six glucopyranoseunits; β-cyclodextrin contains seven glucopyranose units; andγ-cyclodextrin contains eight glucopyranose units. The molecule isbelieved to form a truncated cone having a core opening of 4.7-5.3angstroms, 6.0-6.5 angstroms, and 7.5-8.3 angstroms in α-, β-, orγ-cyclodextrin respectively. The composition according to the inventionmay comprise a mixture of two or more of the α-, β-, or γ-cyclodextrins.Typically, however, the composition according to the invention willcomprise only one of the α-, β-, or γ-cyclodextrins.

Most preferred cyclodextrins in the compositions according to theinvention are amorphous cyclodextrin compounds. By amorphouscyclodextrin is meant non-crystalline mixtures of cyclodextrins whereinthe mixture is prepared from α-, β-, or γ-cyclodextrin. In general, theamorphous cyclodextrin is prepared by non-selective alkylation of thedesired cyclodextrin species. Suitable alkylation agents for thispurpose include but are not limited to propylene oxide, glycidol,iodoacetamide, chloroacetate, and 2-diethylaminoethlychloride. Reactionsare carried out to yield mixtures containing a plurality of componentsthereby preventing crystallization of the cyclodextrin. Variousalkylated cyclodextrins can be made and of course will vary, dependingupon the starting species of cyclodextrin and the alkylating agent used.Among the amorphous cyclodextrins suitable for compositions according tothe invention are hydroxypropyl, hydroxyethyl, glucosyl, maltosyl andmaltotriosyl derivatives of β-cyclodextrin,carboxyamidomethyl-β-cyclodextrin, carboxymethyl-β-cyclodextrin,hydroxypropyl-β-cyclodextrin and diethylamino-β-cyclodextrin.

One example of resveratrol dissolved in the presence of a cyclodextrinis provided in Marier et al., J. Pharmacol. Exp. Therap. 302:369-373(2002), the contents of which are incorporated herein by reference,where a 6 mg/mL solution of resveratrol was prepared using 0.9% salinecontaining 20% hydroxylpropyl-β-cyclodextrin.

As mentioned above, the compositions of matter of the invention comprisean aqueous preparation of preferably substituted amorphous cyclodextrinand one or more sirtuin modulators. The relative amounts of sirtuinmodulators and cyclodextrin will vary depending upon the relative amountof each of the sirtuin modulators and the effect of the cyclodextrin onthe compound. In general, the ratio of the weight of compound of thesirtuin modulators to the weight of cyclodextrin compound will be in arange between 1:1 and 1:100. A weight to weight ratio in a range of 1:5to 1:50 and more preferably in a range of 1:10 to 1:20 of the compoundselected from sirtuin modulators to cyclodextrin are believed to be themost effective for increased circulating availability of the sirtuinmodulator.

Importantly, if the aqueous solution comprising the sirtuin modulatorsand a cyclodextrin is to be administered parenterally, especially viathe intravenous route, a cyclodextrin will be substantially free ofpyrogenic contaminants. Various forms of cyclodextrin, such as forms ofamorphous cyclodextrin, may be purchased from a number of vendorsincluding Sigma-Aldrich, Inc. (St. Louis, Mo., USA). A method for theproduction of hydroxypropyl-β-cyclodextrin is disclosed in Pitha et al.,U.S. Pat. No. 4,727,064 which is incorporated herein by reference.

Additional description of the use of cyclodextrin for solubilizingcompounds can be found in US 2005/0026849, the contents of which areincorporated herein by reference.

Rapidly disintegrating or dissolving dosage forms are useful for therapid absorption, particularly buccal and sublingual absorption, ofpharmaceutically active agents. Fast melt dosage forms are beneficial topatients, such as aged and pediatric patients, who have difficulty inswallowing typical solid dosage forms, such as caplets and tablets.Additionally, fast melt dosage forms circumvent drawbacks associatedwith, for example, chewable dosage forms, wherein the length of time anactive agent remains in a patient's mouth plays an important role indetermining the amount of taste masking and the extent to which apatient may object to throat grittiness of the active agent.

To overcome such problems manufacturers have developed a number of fastmelt solid dose oral formulations. These are available frommanufacturers including Cima Labs, Fuisz Technologies Ltd., Prographarm,R. P. Scherer, Yamanouchi-Shaklee, and McNeil-PPC, Inc. All of thesemanufacturers market different types of rapidly dissolving solid oraldosage forms. See e.g., patents and publications by Cima Labs such asU.S. Pat. Nos. 5,607,697, 5,503,846, 5,223,264, 5,401,513, 5,219,574,and 5,178,878, WO 98/46215, WO 98/14179; patents to Fuisz Technologies,now part of BioVail, such as U.S. Pat. Nos. 5,871,781, 5,869,098,5,866,163, 5,851,553, 5,622,719, 5,567,439, and 5,587,172; U.S. Pat. No.5,464,632 to Prographarm; patents to R. P. Scherer such as U.S. Pat.Nos. 4,642,903, 5,188,825, 5,631,023 and 5,827,541; patents toYamanouchi-Shaklee such as U.S. Pat. Nos. 5,576,014 and 5,446,464;patents to Janssen such as U.S. Pat. Nos. 5,807,576, 5,635,210,5,595,761, 5,587,180 and 5,776,491; U.S. Pat. Nos. 5,639,475 and5,709,886 to Eurand America, Inc.; U.S. Pat. Nos. 5,807,578 and5,807,577 to L.A.B. Pharmaceutical Research; patents to ScheringCorporation such as U.S. Pat. Nos. 5,112,616 and 5,073,374; U.S. Pat.No. 4,616,047 to Laboratoire L. LaFon; U.S. Pat. No. 5,501,861 to TakedaChemicals Inc., Ltd.; and U.S. Pat. No. 6,316,029 to Elan.

In one example of fast melt tablet preparation, granules for fast melttablets made by either the spray drying or pre-compacting processes aremixed with excipients and compressed into tablets using conventionaltablet making machinery. The granules can be combined with a variety ofcarriers including low density, high moldability saccharides, lowmoldability saccharides, polyol combinations, and then directlycompressed into a tablet that exhibits an improved dissolution anddisintegration profile.

The tablets according to the present invention typically have a hardnessof about 2 to about 6 Strong-Cobb units (scu). Tablets within thishardness range disintegrate or dissolve rapidly when chewed.Additionally, the tablets rapidly disentegrate in water. On average, atypical 1.1 to 1.5 gram tablet disintegrates in 1-3 minutes withoutstirring. This rapid disintegration facilitates delivery of the activematerial.

The granules used to make the tablets can be, for example, mixtures oflow density alkali earth metal salts or carbohydrates. For example, amixture of alkali earth metal salts includes a combination of calciumcarbonate and magnesium hydroxide. Similarly, a fast melt tablet can beprepared according to the methods of the present invention thatincorporates the use of A) spray dried extra light calciumcarbonate/maltodextrin, B) magnesium hydroxide and C) a eutectic polyolcombination including Sorbitol Instant, xylitol and mannitol. Thesematerials have been combined to produce a low density tablet thatdissolves very readily and promotes the fast disintegration of theactive ingredient. Additionally, the pre-compacted and spray driedgranules can be combined in the same tablet.

For fast melt tablet preparation, a sirtuin modulator useful in thepresent invention can be in a form such as solid, particulate, granular,crystalline, oily or solution. The sirtuin modulator for use in thepresent invention may be a spray dried product or an adsorbate that hasbeen pre-compacted to a harder granular form that reduces the medicamenttaste. A pharmaceutical active ingredient for use in the presentinvention may be spray dried with a carrier that prevents the activeingredient from being easily extracted from the tablet when chewed.

In addition to being directly added to the tablets of the presentinvention, the medicament drug itself can be processed by thepre-compaction process to achieve an increased density prior to beingincorporated into the formulation.

The pre-compaction process used in the present invention can be used todeliver poorly soluble pharmaceutical materials so as to improve therelease of such pharmaceutical materials over traditional dosage forms.This could allow for the use of lower dosage levels to deliverequivalent bioavailable levels of drug and thereby lower toxicity levelsof both currently marketed drug and new chemical entities. Poorlysoluble pharmaceutical materials can be used in the form ofnanoparticles, which are nanometer-sized particles.

In addition to the active ingredient and the granules prepared from lowdensity alkali earth metal salts and/or water soluble carbohydrates, thefast melt tablets can be formulated using conventional carriers orexcipients and well established pharmaceutical techniques. Conventionalcarriers or excipients include, but are not limited to, diluents,binders, adhesives (i.e., cellulose derivatives and acrylicderivatives), lubricants (i.e., magnesium or calcium stearate, vegetableoils, polyethylene glycols, talc, sodium lauryl sulphate, polyoxyethylene monostearate), disintegrants, colorants, flavorings,preservatives, sweeteners and miscellaneous materials such as buffersand adsorbents.

Additional description of the preparation of fast melt tablets can befound, for example, in U.S. Pat. No. 5,939,091, the contents of whichare incorporated herein by reference.

Pharmaceutical compositions (including cosmetic preparations) maycomprise from about 0.00001 to 100% such as from 0.001 to 10% or from0.1% to 5% by weight of one or more sirtuin-modulating compoundsdescribed herein.

In one embodiment, a sirtuin-modulating compound described herein, isincorporated into a topical formulation containing a topical carrierthat is generally suited to topical drug administration and comprisingany such material known in the art. The topical carrier may be selectedso as to provide the composition in the desired form, e.g., as anointment, lotion, cream, microemulsion, gel, oil, solution, or the like,and may be comprised of a material of either naturally occurring orsynthetic origin. It is preferable that the selected carrier notadversely affect the active agent or other components of the topicalformulation. Examples of suitable topical carriers for use hereininclude water, alcohols and other nontoxic organic solvents, glycerin,mineral oil, silicone, petroleum jelly, lanolin, fatty acids, vegetableoils, parabens, waxes, and the like.

Formulations may be colorless, odorless ointments, lotions, creams,microemulsions and gels.

Sirtuin-modulating compounds may be incorporated into ointments, whichgenerally are semisolid preparations which are typically based onpetrolatum or other petroleum derivatives. The specific ointment base tobe used, as will be appreciated by those skilled in the art, is one thatwill provide for optimum drug delivery, and, preferably, will providefor other desired characteristics as well, e.g., emolliency or the like.As with other carriers or vehicles, an ointment base should be inert,stable, nonirritating and nonsensitizing. As explained in Remington's(supra) ointment bases may be grouped in four classes: oleaginous bases;emulsifiable bases; emulsion bases; and water-soluble bases. Oleaginousointment bases include, for example, vegetable oils, fats obtained fromanimals, and semisolid hydrocarbons obtained from petroleum.Emulsifiable ointment bases, also known as absorbent ointment bases,contain little or no water and include, for example, hydroxystearinsulfate, anhydrous lanolin and hydrophilic petrolatum. Emulsion ointmentbases are either water-in-oil (W/O) emulsions or oil-in-water (O/W)emulsions, and include, for example, cetyl alcohol, glycerylmonostearate, lanolin and stearic acid. Exemplary water-soluble ointmentbases are prepared from polyethylene glycols (PEGs) of varying molecularweight; again, reference may be had to Remington's, supra, for furtherinformation.

Sirtuin-modulating compounds may be incorporated into lotions, whichgenerally are preparations to be applied to the skin surface withoutfriction, and are typically liquid or semiliquid preparations in whichsolid particles, including the active agent, are present in a water oralcohol base. Lotions are usually suspensions of solids, and maycomprise a liquid oily emulsion of the oil-in-water type. Lotions arepreferred formulations for treating large body areas, because of theease of applying a more fluid composition. It is generally necessarythat the insoluble matter in a lotion be finely divided. Lotions willtypically contain suspending agents to produce better dispersions aswell as compounds useful for localizing and holding the active agent incontact with the skin, e.g., methylcellulose, sodiumcarboxymethylcellulose, or the like. An exemplary lotion formulation foruse in conjunction with the present method contains propylene glycolmixed with a hydrophilic petrolatum such as that which may be obtainedunder the trademark Aquaphor™ from Beiersdorf, Inc. (Norwalk, Conn.).

Sirtuin-modulating compounds may be incorporated into creams, whichgenerally are viscous liquid or semisolid emulsions, either oil-in-wateror water-in-oil. Cream bases are water-washable, and contain an oilphase, an emulsifier and an aqueous phase. The oil phase is generallycomprised of petrolatum and a fatty alcohol such as cetyl or stearylalcohol; the aqueous phase usually, although not necessarily, exceedsthe oil phase in volume, and generally contains a humectant. Theemulsifier in a cream formulation, as explained in Remington's, supra,is generally a nonionic, anionic, cationic or amphoteric surfactant.

Sirtuin-modulating compounds may be incorporated into microemulsions,which generally are thermodynamically stable, isotropically cleardispersions of two immiscible liquids, such as oil and water, stabilizedby an interfacial film of surfactant molecules (Encyclopedia ofPharmaceutical Technology (New York: Marcel Dekker, 1992), volume 9).For the preparation of microemulsions, surfactant (emulsifier),co-surfactant (co-emulsifier), an oil phase and a water phase arenecessary. Suitable surfactants include any surfactants that are usefulin the preparation of emulsions, e.g., emulsifiers that are typicallyused in the preparation of creams. The co-surfactant (or “co-emulsifer”)is generally selected from the group of polyglycerol derivatives,glycerol derivatives and fatty alcohols. Preferredemulsifier/co-emulsifier combinations are generally although notnecessarily selected from the group consisting of: glyceryl monostearateand polyoxyethylene stearate; polyethylene glycol and ethylene glycolpalmitostearate; and caprilic and capric triglycerides and oleoylmacrogolglycerides. The water phase includes not only water but also,typically, buffers, glucose, propylene glycol, polyethylene glycols,preferably lower molecular weight polyethylene glycols (e.g., PEG 300and PEG 400), and/or glycerol, and the like, while the oil phase willgenerally comprise, for example, fatty acid esters, modified vegetableoils, silicone oils, mixtures of mono- di- and triglycerides, mono- anddi-esters of PEG (e.g., oleoyl macrogol glycerides), etc.

Sirtuin-modulating compounds may be incorporated into gel formulations,which generally are semisolid systems consisting of either suspensionsmade up of small inorganic particles (two-phase systems) or largeorganic molecules distributed substantially uniformly throughout acarrier liquid (single phase gels). Single phase gels can be made, forexample, by combining the active agent, a carrier liquid and a suitablegelling agent such as tragacanth (at 2 to 5%), sodium alginate (at2-10%), gelatin (at 2-15%), methylcellulose (at 3-5%), sodiumcarboxymethylcellulose (at 2-5%), carbomer (at 0.3-5%) or polyvinylalcohol (at 10-20%) together and mixing until a characteristic semisolidproduct is produced. Other suitable gelling agents includemethylhydroxycellulose, polyoxyethylene-polyoxypropylene,hydroxyethylcellulose and gelatin. Although gels commonly employ aqueouscarrier liquid, alcohols and oils can be used as the carrier liquid aswell.

Various additives, known to those skilled in the art, may be included informulations, e.g., topical formulations. Examples of additives include,but are not limited to, solubilizers, skin permeation enhancers,opacifiers, preservatives (e.g., anti-oxidants), gelling agents,buffering agents, surfactants (particularly nonionic and amphotericsurfactants), emulsifiers, emollients, thickening agents, stabilizers,humectants, colorants, fragrance, and the like. Inclusion ofsolubilizers and/or skin permeation enhancers is particularly preferred,along with emulsifiers, emollients and preservatives. An optimum topicalformulation comprises approximately: 2 wt. % to 60 wt. %, preferably 2wt. % to 50 wt. %, solubilizer and/or skin permeation enhancer; 2 wt. %to 50 wt. %, preferably 2 wt. % to 20 wt. %, emulsifiers; 2 wt. % to 20wt. % emollient; and 0.01 to 0.2 wt. % preservative, with the activeagent and carrier (e.g., water) making of the remainder of theformulation.

A skin permeation enhancer serves to facilitate passage of therapeuticlevels of active agent to pass through a reasonably sized area ofunbroken skin. Suitable enhancers are well known in the art and include,for example: lower alkanols such as methanol ethanol and 2-propanol;alkyl methyl sulfoxides such as dimethylsulfoxide (DMSO),decylmethylsulfoxide (C₁₀ MSO) and tetradecylmethyl sulfboxide;pyrrolidones such as 2-pyrrolidone, N-methyl-2-pyrrolidone andN-(hydroxyethyl)pyrrolidone; urea; N,N-diethyl-m-toluamide; C₂-C₆alkanediols; miscellaneous solvents such as dimethyl formamide (DMF),N,N-dimethylacetamide (DMA) and tetrahydrofurfuryl alcohol; and the1-substituted azacycloheptan-2-ones, particularly1-n-dodecylcyclazacycloheptan-2-one (laurocapram; available under thetrademark Azone® from Whitby Research Incorporated, Richmond, Va.).

Examples of solubilizers include, but are not limited to, the following:hydrophilic ethers such as diethylene glycol monoethyl ether(ethoxydiglycol, available commercially as Transcutol®) and diethyleneglycol monoethyl ether oleate (available commercially as Softcutol®);polyethylene castor oil derivatives such as polyoxy 35 castor oil,polyoxy 40 hydrogenated castor oil, etc.; polyethylene glycol,particularly lower molecular weight polyethylene glycols such as PEG 300and PEG 400, and polyethylene glycol derivatives such as PEG-8caprylic/capric glycerides (available commercially as Labrasol®); alkylmethyl sulfoxides such as DMSO; pyrrolidones such as 2-pyrrolidone andN-methyl-2-pyrrolidone; and DMA. Many solubilizers can also act asabsorption enhancers. A single solubilizer may be incorporated into theformulation, or a mixture of solubilizers may be incorporated therein.

Suitable emulsifiers and co-emulsifiers include, without limitation,those emulsifiers and co-emulsifiers described with respect tomicroemulsion formulations. Emollients include, for example, propyleneglycol, glycerol, isopropyl myristate, polypropylene glycol-2 (PPG-2)myristyl ether propionate, and the like.

Other active agents may also be included in formulations, e.g., otheranti-inflammatory agents, analgesics, antimicrobial agents, antifungalagents, antibiotics, vitamins, antioxidants, and sunblock agentscommonly found in sunscreen formulations including, but not limited to,anthranilates, benzophenones (particularly benzophenone-3), camphorderivatives, cinnamates (e.g., octyl methoxycinnamate), dibenzoylmethanes (e.g., butyl methoxydibenzoyl methane), p-aminobenzoic acid(PABA) and derivatives thereof, and salicylates (e.g., octylsalicylate).

In certain topical formulations, the active agent is present in anamount in the range of approximately 0.25 wt. % to 75 wt. % of theformulation, preferably in the range of approximately 0.25 wt. % to 30wt. % of the formulation, more preferably in the range of approximately0.5 wt. % to 15 wt. % of the formulation, and most preferably in therange of approximately 1.0 wt. % to 10 wt. % of the formulation.

Topical skin treatment compositions can be packaged in a suitablecontainer to suit its viscosity and intended use by the consumer. Forexample, a lotion or cream can be packaged in a bottle or a roll-ballapplicator, or a propellant-driven aerosol device or a container fittedwith a pump suitable for finger operation. When the composition is acream, it can simply be stored in a non-deformable bottle or squeezecontainer, such as a tube or a lidded jar. The composition may also beincluded in capsules such as those described in U.S. Pat. No. 5,063,507.Accordingly, also provided are closed containers containing acosmetically acceptable composition as herein defined.

In an alternative embodiment, a pharmaceutical formulation is providedfor oral or parenteral administration, in which case the formulation maycomprises a modulating compound-containing microemulsion as describedabove, but may contain alternative pharmaceutically acceptable carriers,vehicles, additives, etc. particularly suited to oral or parenteral drugadministration. Alternatively, a modulating compound-containingmicroemulsion may be administered orally or parenterally substantiallyas described above, without modification.

Phospholipids complexes, e.g., resveratrol-phospholipid complexes, andtheir preparation are described in U.S. Patent Application PublicationNo. 2004/116386. Methods for stabilizing active components usingpolyol/polymer microcapsules, and their preparation are described inUS20040108608. Processes for dissolving lipophilic compounds in aqueoussolution with amphiphilic block copolymers are described in WO04/035013.

Conditions of the eye can be treated or prevented by, e.g., systemic,topical, intraocular injection of a sirtuin-modulating compound, or byinsertion of a sustained release device that releases asirtuin-modulating compound. A sirtuin-modulating compound thatincreases or decreases the level and/or activity of a sirtuin proteinmay be delivered in a pharmaceutically acceptable ophthalmic vehicle,such that the compound is maintained in contact with the ocular surfacefor a sufficient time period to allow the compound to penetrate thecorneal and internal regions of the eye, as for example the anteriorchamber, posterior chamber, vitreous body, aqueous humor, vitreoushumor, cornea, iris/ciliary, lens, choroid/retina and sclera. Thepharmaceutically-acceptable ophthalmic vehicle may, for example, be anointment, vegetable oil or an encapsulating material. Alternatively, thecompounds of the invention may be injected directly into the vitreousand aqueous humour. In a further alternative, the compounds may beadministered systemically, such as by intravenous infusion or injection,for treatment of the eye.

Sirtuin-modulating compounds described herein may be stored in oxygenfree environment according to methods in the art. For example,resveratrol or analog thereof can be prepared in an airtight capsule fororal administration, such as Capsugel from Pfizer, Inc.

Cells, e.g., treated ex vivo with a sirtuin-modulating compound, can beadministered according to methods for administering a graft to asubject, which may be accompanied, e.g., by administration of animmunosuppressant drug, e.g., cyclosporin A. For general principles inmedicinal formulation, the reader is referred to Cell Therapy: Stem CellTransplantation, Gene Therapy, and Cellular Immunotherapy, by G. Morstyn& W. Sheridan eds, Cambridge University Press, 1996; and HematopoieticStem Cell Therapy, E. D. Ball, J. Lister & P. Law, ChurchillLivingstone, 2000.

Toxicity and therapeutic efficacy of sirtuin-modulating compounds can bedetermined by standard pharmaceutical procedures in cell cultures orexperimental animals. The LD₅₀ is the dose lethal to 50% of thepopulation. The ED50 is the dose therapeutically effective in 50% of thepopulation. The dose ratio between toxic and therapeutic effects(LD₅₀/ED₅₀) is the therapeutic index. Sirtuin-modulating compounds thatexhibit large therapeutic indexes are preferred. Whilesirtuin-modulating compounds that exhibit toxic side effects may beused, care should be taken to design a delivery system that targets suchcompounds to the site of affected tissue in order to minimize potentialdamage to uninfected cells and, thereby, reduce side effects.

The data obtained from the cell culture assays and animal studies can beused in formulating a range of dosage for use in humans. The dosage ofsuch compounds may lie within a range of circulating concentrations thatinclude the ED50 with little or no toxicity. The dosage may vary withinthis range depending upon the dosage form employed and the route ofadministration utilized. For any compound, the therapeutically effectivedose can be estimated initially from cell culture assays. A dose may beformulated in animal models to achieve a circulating plasmaconcentration range that includes the IC₅₀ (i.e., the concentration ofthe test compound that achieves a half-maximal inhibition of symptoms)as determined in cell culture. Such information can be used to moreaccurately determine useful doses in humans. Levels in plasma may bemeasured, for example, by high performance liquid chromatography.

6. Kits

Also provided herein are kits, e.g., kits for therapeutic purposes orkits for modulating the lifespan of cells or modulating apoptosis. A kitmay comprise one or more sirtuin-modulating compounds, e.g., inpremeasured doses. A kit may optionally comprise devices for contactingcells with the compounds and instructions for use. Devices includesyringes, stents and other devices for introducing a sirtuin-modulatingcompound into a subject (e.g., the blood vessel of a subject) orapplying it to the skin of a subject.

Another type of kit contemplated by the invention are kits foridentifying sirtuin-modulating compounds. Such kits contain (1) asirtuin or sirtuin-containing material and (2) a sirtuin-modulatingcompound of the invention, which are in separate vessels. Such kits canbe used, for example, to perform a competition-type assay to test othercompounds (typically provided by the user) for sirtuin-modulatingactivity. In certain embodiments, these kits further comprise means fordetermining sirtuin activity (e.g., a peptide with an appropriateindicator, such as those disclosed in the Exemplification).

In yet another embodiment, the invention provides a composition ofmatter comprising a sirtruin modulator of this invention and anothertherapeutic agent [the same ones used in combination therapies andcombination compositions] in separate dosage forms, but associated withone another. The term “associated with one another” as used herein meansthat the separate dosage forms are packaged together or otherwiseattached to one another such that it is readily apparent that theseparate dosage forms are intended to be sold and administered as partof the same regimen. The agent and the sirtruin modulator are preferablypackaged together in a blister pack or other multi-chamber package, oras connected, separately sealed containers (such as foil pouches or thelike) that can be separated by the user (e.g., by tearing on score linesbetween the two containers).

In still another embodiment, the invention provides a kit comprising inseparate vessels, a) a sirtruin modulator of this invention; and b)another another therapeutic agent such as those described elsewhere inthe specification.

The practice of the present methods will employ, unless otherwiseindicated, conventional techniques of cell biology, cell culture,molecular biology, transgenic biology, microbiology, recombinant DNA,and immunology, which are within the skill of the art. Such techniquesare explained fully in the literature. See, for example, MolecularCloning A Laboratory Manual, 2^(nd) Ed., ed. by Sambrook, Fritsch andManiatis (Cold Spring Harbor Laboratory Press: 1989); DNA Cloning,Volumes I and II (D. N. Glover ed., 1985); Oligonucleotide Synthesis (M.J. Gait ed., 1984); Mullis et al. U.S. Pat. No. 4,683,195; Nucleic AcidHybridization (B. D. Hames & S. J. Higgins eds. 1984); Transcription AndTranslation (B. D. Hames & S. J. Higgins eds. 1984); Culture Of AnimalCells (R. I. Freshney, Alan R. Liss, Inc., 1987); Immobilized Cells AndEnzymes (IRL Press, 1986); B. Perbal, A Practical Guide To MolecularCloning (1984); the treatise, Methods In Enzymology (Academic Press,Inc., N.Y.); Gene Transfer Vectors For Mammalian Cells (J. H. Miller andM. P. Calos eds., 1987, Cold Spring Harbor Laboratory); Methods InEnzymology, Vols. 154 and 155 (Wu et al. eds.), Immunochemical MethodsIn Cell And Molecular Biology (Mayer and Walker, eds., Academic Press,London, 1987); Handbook Of Experimental Immunology, Volumes I-IV (D. M.Weir and C. C. Blackwell, eds., 1986); Manipulating the Mouse Embryo,(Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1986).

EXEMPLIFICATION

The invention now being generally described, it will be more readilyunderstood by reference to the following examples which are includedmerely for purposes of illustration of certain aspects and embodimentsof the present invention, and are not intended to limit the invention inany way.

Example 1 Synthesis and Characterization of Sirtuin Modulators

General Schemes:

Experimental Section:Abbreviations Used in Experimental Section:

-   HATU=O-(7-Azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium    hexafluorophosphate-   NMM=4-Methylmorpholine-   DIEA=N,N-Diisopropylethylamine-   DMF=N,N-Dimethylformamide-   CH₂Cl₂=Dichloromethane-   EtOAc=Ethyl acetate-   MeOH=Methanol-   Na₂SO₄=Sodium sulfate-   PPA=Polyphosphoric acid-   Et₃N=Triethylamine-   rt=room temperature

Preparation of 3-(thiazolo[5,4-c]pyridin-2-yl)benzenamine

4-aminopyridin-3-yl diisopropylcarbamodithioate was prepared accordingto the procedures outlined in Smith et al, Sulfur Lett. 1994 vol 17, p.197 and E. Ma, Molecules 2003, vol 8, p. 678-686.

220 mg of 4-aminopyridin-3-yl diisopropylcarbamodithioate (0.81 mmol)was dissolved in 6 mL of methylene chloride and cooled to 10 degrees C.(ice bath) along with triethylamine (0.175 mL, 1.5 eq). 3-Nitrobenzoylchloride (150 mg, 1 eq, 0.81 mmol) was dissolved in 3 mL of methylenechloride and then added to the cooled solution of 4-aminopyridin-3-yldiisopropylcarbamodithioate. The reaction mixture was warmed to rt andstirred for 45 min. The reaction mixture was diluted with 5 mL ofmethylene chloride and washed with brine. The organic layer was dried(Na₂SO₄) and concentrated to afford 280 mg of compound of theintermediate amide (83% crude yield).

This intermediate amide (280 mg) was suspended in 5 mL of 4 N aq HCl andstirred under reflux for 30 min. The reaction mixture was cooled to rtand neutralized with 3 N NaOH and extracted with methylene chloride. Theorganic layer was dried (Na₂SO₄) and concentrated under reduced pressureto afford 200 mg of compound of 2-(3-nitrophenyl)thiazolo[5,4-c]pyridine(95% crude yield).

310 mg of 2-(3-nitrophenyl)thiazolo[5,4-c]pyridine (1.2 mmol) was mixedwith 30 mL of MeOH along with 6 mL of water. Sodium hydrosulfide hydrate(6 eq, 7.24 mmol, 400 mg) was added and the reaction mixture was stirredunder reflux for 3 hours. The reaction mixture was cooled to roomtemperature and concentrated. The aqueous layer was extracted withmethylene chloride. The combined organic layers were dried (Na₂SO₄) andconcentrated to afford 230 mg of3-(thiazolo[5,4-c]pyridin-2-yl)benzenamine (84% crude yield) (MS,M⁺+H=228).

Preparation of Compound 115

In a typical run, 40 mg of 3-(thiazolo[5,4-c]pyridin-2-yl)benzenamine(0.176 mmol) was suspended in 1 mL of pyridine along with 1 eq of3,4,5-trimethoxybenzoyl chloride. The reaction mixture was then heatedin a Biotage microwave reactor at 160 degree for 10 min. It was thencooled to room temperature and concentrated under reduced pressure. Theresulting residue was purified by chromatography using a 9:1 mixture ofCH₂Cl₂ to MeOH (MS, M⁺+H=422)

Preparation of Compounds 113 and 114

The same procedure used in the preparation of Compound 115 was employedusing the appropriate acid chloride.

Preparation of Compound 99

In a typical run, 3-(thiazolo[5,4-c]pyridin-2-yl)benzenamine (25 mg,0.11 mmol) was suspended in 1 mL of pyridine along with 18 mg of3,4-(methylenedioxy)phenyl isocyanate. The reaction mixture was thenheated in a Biotage microwave reactor at 140 degree for 10 min. It wasthen cooled to room temperature and concentrated. The resulting residuewas purified by chromatography using a 9:1 mixture of CH₂Cl₂ to MeOH(MS, M⁺+H=391).

Preparation of methyl 3-amino-5-(thiazolo[5,4-c]pyridin-2-yl)benzoate

Mono-methyl 5-nitro-isophthlate (1.25 g, 5.58 mmol) was suspended in 25mL of CH₂Cl₂ and oxalyl chloride (0.49 mL, 5.58 mmol) was added. After 3drops of DMF was added, the reaction mixture was stirred at roomtemperature until all gas evolution had ceased and all the solids haddissolved. This freshly prepared solution of the acid chloride was thenadded dropwise to a solution of 4-aminopyridin-3-yldiisopropylcarbamodithioate (1.5 g, 5.58 mmol) and triethylamine (0.77mL, 5.58 mmol) in 50 mL of CH₂Cl₂ at 0° C. The resulting reactionmixture was warmed to room temperature and stirred for 1 hour. It wasthen quenched with 25 mL of brine and the two layers were separated. Theorganic layer was dried (Na₂SO₄) and concentrated. The resulting residuewas suspended in 25 mL of 2 N HCl and stirred under reflux for 30 min.It was then cooled to room temperature and the solids were collected byfiltration and dried to afford 1.0 g of methyl3-nitro-5-(thiazolo[5,4-c]pyridin-2-yl)benzoate. This material was thenmixed with 20 mL of methanol and 3 mL of water along with 1 g of sodiumhydrosulfide hydrate and stirred under reflux for 2 hours. The resultingreaction mixture was cooled and concentrated. The aqueous layer wasextracted with CH₂Cl₂. The combined organic layers were dried (Na₂SO₄)and concentrated to afford 150 mg of methyl3-amino-5-(thiazolo[5,4-c]pyridin-2-yl)benzoate (MS, M⁺+H=286).

Preparation of Compound 133

Methyl 3-amino-5-(thiazolo[5,4-c]pyridin-2-yl)benzoate (150 mg, 0.526mmol) was mixed together with 3,4,5-trimethoxybenzoyl chloride (121 mg,0.526 mmol) in 1 mL of pyridine. The reaction mixture was reacted in aBiotage microwave reactor at 160 degrees for 10 min. It was then cooledto room temperature and concentrated. The resulting residue was purifiedby chromatography using a 9:1 mixture of CH₂Cl₂ to MeOH to afford 90 mgof methyl3-(thiazolo[5,4-c]pyridin-2-yl)-5-(3,4,5-trimethoxybenzamido)benzoate(36% yield) (MS, M⁺+H=480).

Preparation of Compound 134

Methyl3-(thiazolo[5,4-c]pyridin-2-yl)-5-(3,4,5-trimethoxybenzamido)benzoate(80 mg, 0.167 mmol) was dissolved in 5 mL of THF and 2 mL of watercontaining 30 mg of sodium hydroxide. The reaction mixture was stirredat room temperature for 1 hour. It was then acidified to pH 4 with 6 NHCl and concentrated. The solids were collected by filtration and driedto afford 60 mg of3-(thiazolo[5,4-c]pyridin-2-yl)-5-(3,4,5-trimethoxybenzamido)benzoicacid (78% yield) (MS, M⁺+H=466).

Preparation of Compound 135

3-(thiazolo[5,4-c]pyridin-2-yl)-5-(3,4,5-trimethoxybenzamido)benzoicacid (50 mg, 0.108 mmol) was suspended in 2 mL of anhydrous THF andcooled to 0° C. along with 1 eq of NMM. Isobutyl chloroformate (1 eq)was added and the reaction mixture was stirred for 45 min. NaBH₄ (1 eq)was then added as a solution in 0.5 mL of water. The reaction mixturewas stirred for 30 min and then warmed to room temperature andconcentrated. The aqueous layer was extracted with CH₂Cl₂. The combinedorganic layers were dried (Na₂SO₄) and concentrated to afford the crudeproduct. This was purified by chromatography using a 9:1 mixture ofCH₂Cl₂ to MeOH (MS, M⁺+H=452).

Preparation of 3-(oxazolo[5,4-b]pyridin-2-yl)benzenamine

2-Chloropyridin-3-amine (3.20 g, 0.025 mol) was suspended in 15 mL ofpyridine and added slowly to a suspension of 3-nitrobenzoyl chloride(4.64 g, 0.025 mol) in 15 mL of pyridine in an ice bath. The reactionmixture was slowly warmed to room temperature and stirred overnight. Thenext day, the reaction mixture was cooled in an ice bath and 30 mL ofglacial acetic acid was added slowly. The resulting mixture was dilutedwith 200 mL of EtOAc and washed with water (3×20 mL). The organic layerwas then dried (Na₂SO₄) and concentrated. The resulting residue wasmixed with 10 mL of PPA and stirred at 160 degree for 6 hours. Thereaction mixture was then poured carefully into 150 mL of water. The pHwas brought to about 5 with solid NaOH and the solids were collected byfiltration and dried. This material was mixed with 50 mL of MeOH andfiltered. The filtrate was concentrated to afford 2.1 g of2-(3-nitrophenyl)oxazolo[5,4-b]pyridine (35% yield) (MS, M⁺+H=242).

2-(3-Nitrophenyl)oxazolo[5,4-b]pyridine (600 mg, 2.49 mmol) was mixedwith 25 mL of MeOH and 4 mL of water along with 837 mg of sodiumhydrosulfide hydrate (14.9 mmol). The reaction mixture was stirred underreflux for 3 hours. It was then cooled to room temperature andconcentrated. The aqueous layer was extracted with CH₂Cl₂. The combinedorganic layers were dried (Na₂SO₄) and concentrated to afford 520 mg of3-(oxazolo[5,4-b]pyridin-2-yl)benzenamine (quantitative crude yield)(MS, M⁺+H=212).

Preparation of Compound 112

3-(Oxazolo[5,4-b]pyridin-2-yl)benzenamine was reacted with3,4-dimethoxybenzoyl chloride under the microwave reaction conditionsdescribed earlier. The crude product was purified by chromatographyusing a 9:1 mixture of CH₂Cl₂ to MeOH (MS, M⁺+H=376)

Preparation of Compound 74 and 111

The same procedure used in the preparation of Compound 112 was employedusing 3,4-dimethoxyphenyl sulfonyl chloride and 3-dimethylaminobenzoylchloride hydrochloride, respectively. The final product was purified bychromatography using a 9:1 mixture of CH₂Cl₂ to MeOH.

Preparation of 3-nitro-5-(oxazolo[4,5-b]pyridin-2-yl)benzoic acid andmethyl 3-nitro-5-(oxazolo[4,5-b]pyridin-2-yl)benzoate

2-Amino-3-hydroxypyridine (1.00 g, 9.16 mmol) was dissolved in 20 mL ofDMF along with 2.06 g of mono-methyl 5-nitroisophthlate (9.16 mmol), 5.2g of HATU (13.7 mmol) and 3.2 mL of DIEA (18.3 mmol). The reactionmixture was stirred at room temperature for 18 hours. It was thendiluted with 200 mL of EtOAc and washed with water (3×25 mL). Theorganic layer was dried (Na₂SO₄) and concentrated. The resulting residuewas mixed with 10 mL of PPA and stirred at 160 degree for 6 hours. Thereaction mixture was then poured carefully into 200 mL of water and thepH was brought to 5 with solid NaOH. The solids were collected byfiltration and dried to afford the product as a 1:1 mixture of themethyl ester and the acid. This mixture was separated by suspending in150 mL of CH₂Cl₂ and then filtered. The filtrate was the methyl ester(MS, M⁺+H=300) and the solids were the desired acid, namely,3-nitro-5-(oxazolo[4,5-b]pyridin-2-yl)benzoic acid (MS, M⁺+H=286).

Preparation of3-amino-N-(2-(dimethylamino)ethyl)-5-(oxazolo[4,5-b]pyridin-2-yl)benzamide

3-Nitro-5-(oxazolo[4,5-b]pyridin-2-yl)benzoic acid (250 mg, 0.877 mmol)was dissolved in 5 mL of DMF along with 1 eq ofN,N-dimethylethylenediamine, 500 mg of HATU (1.5 eq) and 0.3 mL of DIEA(2 eq). The reaction mixture was stirred at room temperature for 18hours. It was then diluted with 50 mL of EtOAc and washed with water.The organic layer was dried (Na₂SO₄) and concentrated to afford thenitro amide intermediate. This was dissolved in 50 mL of MeOH and 5 mLof water along with 200 mg of sodium hydrosulfide hydrate (4 eq). Thereaction mixture was stirred under reflux for 1 hour. It was thenconcentrated to dryness and mixed with 50 mL of 1:1 CH₂Cl₂/MeOH andfiltered. The filtrate was concentrated to afford essentiallyquantitative yield of3-amino-N-(2-(dimethylamino)ethyl)-5-(oxazolo[4,5-b]pyridin-2-yl)benzamide(MS, M⁺+H=326).

Preparation of Compound 153

3-amino-N-(2-(dimethylamino)ethyl)-5-(oxazolo[4,5-b]pyridin-2-yl)benzamidewas reacted with 3,4-dimethoxybenzoyl chloride under the same microwaveconditions as described earlier. The crude product was purified bychromatography using a 9:1 mixture of CH₂Cl₂ to MeOH (MS, M⁺+H=490).

Preparation of Compounds 154 and 155:

The same procedure used in the preparation of Compound 153 was employedusing the appropriate acid chloride.

Preparation of tert-butyl4-(3-amino-5-(oxazolo[4,5-b]pyridin-2-yl)benzoyl)piperazine-1-carboxylate

3-Nitro-5-(oxazolo[4,5-b]pyridin-2-yl)benzoic acid (250 mg, 0.877 mmol)was dissolved in 5 mL of DMF along with 1 eq of Boc-piperazine (163 mg),500 mg of HATU (1.5 eq) and 0.3 mL of DIEA (2 eq). The reaction mixturewas stirred at room temperature for 18 hours. It was then diluted with50 mL of EtOAc and washed with water. The organic layer was dried(Na₂SO₄) and concentrated to afford the nitro amide intermediate. Thiswas dissolved in 50 mL of MeOH and 5 mL of water along with 200 mg ofsodium hydrosulfide hydrate (4 eq). The reaction mixture was stirredunder reflux for 1 hour. The reaction mixture was concentrated and theaqueous layer was extracted with CH₂Cl₂. The combined organic layerswere dried (Na₂SO₄) and concentrated to afford 300 mg of tert-butyl4-(3-amino-5-(oxazolo[4,5-b]pyridin-2-yl)benzoyl)piperazine-1-carboxylate.(MS, M⁺+H=424).

Preparation of Compound 107

tert-Butyl4-(3-amino-5-(oxazolo[4,5-b]pyridin-2-yl)benzoyl)piperazine-1-carboxylate(100 mg, 0.236 mmol) was dissolved in 1 mL of pyridine along with 47 mgof 3,4-dimethoxybenzoyl chloride (1 eq). The reaction mixture was heatedin a Biotage microwave reactor for 10 min. It was then cooled to roomtemperature and concentrated. The resulting residue was purified bychromatography using a 9:1 mixture of CH₂Cl₂ to MeOH to afford 120 mg ofthe Boc-protected diamide derivative. This was treated with 2 mL of 25%TFA in CH₂Cl₂ and allowed to stand at room temperature for 1 hour. Itwas then concentrated and triturated with Et₂O to afford3,4-dimethoxy-N-(3-(oxazolo[4,5-b]pyridin-2-yl)-5-(piperazine-1-carbonyl)phenyl)benzamideas the TFA salt (MS, M⁺+H=488).

Preparation of Compounds 138 and 139:

The same procedure used in the preparation of Compound 107 was employedusing the appropriate acid chloride.

Preparation of Compound 136

Methyl 3-nitro-5-(oxazolo[4,5-b]pyridin-2-yl)benzoate (2.70 g) was mixedwith 100 mL of MeOH and 20 mL of water along with 3 g of sodiumhydrosulfide hydrate (6 eq). The reaction mixture was stirred underreflux for 1 hour. It was then cooled to room temperature andconcentrated. The aqueous layer was extracted with CH₂Cl₂. The combinedorganic layers were dried (Na₂SO₄) and concentrated to afford 600 mg ofthe intermediate amine. A portion of this intermediate amine (50 mg) wasreacted with 1 eq of 3,4-dimethoxybenzoyl chloride under the samemicrowave reaction conditions described earlier. The crude product waspurified by chromatography using a 9:1 mixture of CH₂Cl₂ to MeOH (MS,M⁺+H=434).

Preparation of Compound 137

Methyl 3-(3,4-dimethoxybenzamido)-5-(oxazolo[4,5-b]pyridin-2-yl)benzoate(20 mg) was dissolved in 2 mL of THF and 1 mL of water containing 2 eqof NaOH. The reaction mixture was stirred at room temperature for 1hour. It was then acidified to pH 5 with 6 N HCl and concentrated. Theresulting solids were collected by filtration and dried to afford theacid (MS, M⁺+H=420).

Preparation of Compounds 79, 80 and 81:

3-(2-Methylthiazol-4-yl)benzenamine (Aldrich) was subjected to the samemicrowave reaction conditions described earlier using the appropriateacid chloride. The final products were purified by chromatography usinga 9:1 mixture of CH₂Cl₂ to MeOH.

Preparation of 2-amino-3-hydroxy-6-methylpyridine

7.00 g of 3-hydroxy-6-methyl-2-nitropyridine (Aldrich, 0.045 mol) wasdissolved in 250 mL of MeOH and 20 mL of H2O along with 15 g of sodiumhydrosulfide hydrate (6 eq, 0.27 mol). The reaction mixture was stirredunder reflux for 5 hours. The reaction mixture was cooled to roomtemperature and diluted with 300 mL of absolute EtOH and concentrated todryness. The resulting residue was dissolved in 300 mL of CH₂Cl₂ and 10mL of MeOH. The mixture was sonicated for 10 min and allowed to stand atroom temperature for 3 hours. The resulting salts that precipitated outat this point were removed by filtration. The filtrate was concentratedto afford the crude amine. This was purified by passing through a shortplug of silica gel and eluting with 95% CH₂Cl₂, 4% MeOH and 1% Et₃N. Atotal of 5.0 g of the desired amine was obtained (89% yield).

Preparation of 5-methyl-2-(3-nitro-phenyl)-oxazolo[4,5-b]pyridine

1.20 g of 2-amino-3-hydroxy-6-methylpyridine (9.68 mmol) was dissolvedin 20 mL of DMF along with 1.60 g of 3-nitrobenzoic acid (1 eq), 4.4 gof HATU (Novabiochem, 1.2 eq. 11.6 mmol) and 2.5 mL of DIEA (1.5 eq,14.5 mmol). The reaction mixture was stirred at room temperature for 18hrs. It was then diluted with 250 mL of EtOAc and washed with water(4×15 mL). The organic layer was dried (Na₂SO₄) and concentrated toafford the crude product. Purification by chromatography (Isco, gradientelution, pentane to 80% EtOAc/pentane) afforded 1.10 g of the desiredamide intermediate (42% yield).

This amide intermediate (1.10 g, 4.00 mmol) was mixed with 7 mL of PPAand stirred at 150° C. for 5 hrs. The reaction mixture was cooled toabout 80° C. and diluted with 200 mL of water. This mixture was cooledin an ice bath and the pH was slowly brought to 5 using solid NaOH. Theresulting solids were collected by filtration and dried to afford 600 mgof the desired product, namely,5-methyl-2-(3-nitro-phenyl)-oxazolo[4,5-b]pyridine (24% overall yieldfor the 2 steps). LC/MS showed >95% purity.

Preparation of 5-bromomethyl-2-(3-nitro-phenyl)-oxazolo[4,5-b]pyridine

In a typical run, 400 mg of5-methyl-2-(3-nitro-phenyl)-oxazolo[4,5-b]pyridine (1.57 mmol) wassuspended in 50 mL of CCl₄ along with 280 mg (1.57 mmol) of NBS and 20mg of benzoyl peroxide. The reaction mixture was stirred under refluxfor 4 hours. LC/MS showed about 50% conversion to the desired bromide.After another 2 hours of reflux, no additional change was observed.Another 1 eq of NBS was added and reflux was continued for another 4hours. LC/MS showed complete conversion. The reaction mixture wasconcentrated and the resulting crude product was purified bychromatography (Isco, gradient elution, CH₂Cl₂ to 9:1 CH₂Cl₂/MeOH) toafford essentially quantitative yield of5-bromomethyl-2-(3-nitro-phenyl)-oxazolo[4,5-b]pyridine.

Preparation of4-[2-(3-nitro-phenyl)-oxazolo[4,5-b]pyridin-5-ylmethyl]-piperazine-1-carboxylicacid tert-butyl ester

5-Bromomethyl-2-(3-nitro-phenyl)-oxazolo[4,5-b]pyridine (250 mg, 0.75mmol) was dissolved in 5 mL of CH₃CN along with Et₃N (0.200 mL, 1.5mmol) and 140 mg of Boc-piperazine. The reaction mixture was stirred atroom temperature for 18 hours. It was then concentrated and theresulting residue was mixed with 25 mL of CH₂Cl₂ and washed with brine.The organic layer was dried (Na₂SO₄) and concentrated to affordessentially quantitative yield of4-[2-(3-nitro-phenyl)-oxazolo[4,5-b]pyridin-5-ylmethyl]-piperazine-1-carboxylicacid tert-butyl ester.

Preparation of4-[2-(3-amino-phenyl)-oxazolo[4,5-b]pyridin-5-ylmethyl]-piperazine-1-carboxylicacid tert-butyl ester

4-[2-(3-Nitro-phenyl)-oxazolo[4,5-b]pyridin-5-ylmethyl]-piperazine-1-carboxylicacid tert-butyl ester (330 mg, 0.75 mmol) was dissolved in 6 mL of MeOHand 2 mL of water along with 210 mg of sodium hydrosulfide hydrate (3.75mmol). The reaction mixture was stirred under reflux for 4 hours. Thereaction appeared to be complete based on LC/MS. The reaction mixturewas cooled to room temperature and concentrated. The aqueous layer wasextracted with CH₂Cl₂. The combined organic layers were dried (Na₂SO₄)and concentrated to afford essentially quantitative yield of4-[2-(3-amino-phenyl)-oxazolo[4,5-b]pyridin-5-ylmethyl]-piperazine-1-carboxylicacid tert-butyl ester.

Preparation of Compound 166

4-[2-(3-Amino-phenyl)-oxazolo[4,5-b]pyridin-5-ylmethyl]-piperazine-1-carboxylicacid tert-butyl ester (83 mg, 0.2 mmol) was dissolved in 1 mL ofpyridine along with 1 eq of 3-dimethylamino benzoyl chloridehydrochloride (44 mg, 0.2 mmol). The reaction mixture was heated in aBiotage microwave reactor (160° C.) for 10 min. It was then cooled toroom temperature and concentrated. The resulting residue was purified bychromatography (Isco, gradient elution, CH₂Cl₂ to 95% CH₂Cl₂, 4% MeOHand 1% Et₃N) to afford 25 mg of4-{2-[3-(3-dimethylamino-benzoylamino)-phenyl]-oxazolo[4,5-b]pyridin-5-ylmethyl}-piperazine-1-carboxylicacid tert-butyl ester (23% yield).

4-{2-[3-(3-Dimethylamino-benzoylamino)-phenyl]-oxazolo[4,5-b]pyridin-5-ylmethyl}-piperazine-1-carboxylicacid tert-butyl ester (25 mg, 0.045 mmol) was dissolved in 1 mL of 25%TFA in CH₂Cl₂ and allowed to stand at room temperature for 1 hour. Itwas then concentrated and the resulting residue was triturated with Et₂Oto afford essentially quantitative yield of3-dimethylamino-N-[3-(5-piperazin-1-ylmethyl-oxazolo[4,5-b]pyridin-2-yl)-phenyl]-benzamideas the TFA salt.

Preparation of Compound 514:

The preparation followed essentially the same procedure as detailed inthe preparation of Compound 166 except that 2-quinoxaloyl chloride wasused as the acid chloride component.

Preparation of 3-nitro-5-(oxazolo[4,5-b]pyridin-2-yl)benzoic acid andmethyl 3-nitro-5-(oxazolo[4,5-b]pyridin-2-yl)benzoate

2-Amino-3-hydroxypyridine (1.00 g, 9.16 mmol) was dissolved in 20 mL ofDMF along with 2.06 g of mono-methyl 5-nitroisophthlate (9.16 mmol), 5.2g of HATU (13.7 mmol) and 3.2 mL of DIEA (18.3 mmol). The reactionmixture was stirred at room temperature for 18 hours. It was thendiluted with 200 mL of EtOAc and washed with water (3×25 mL). Theorganic layer was dried (Na₂SO₄) and concentrated. The resulting residuewas mixed with 10 mL of PPA and stirred at 160 degree for 6 hours. Thereaction mixture was then poured carefully into 200 mL of water and thepH was brought to 5 with solid NaOH. The solids were collected byfiltration and dried to afford the product as a 1:1 mixture of themethyl ester and the acid. This mixture was separated by suspending itin 50 mL of CH₂Cl₂ and then filtering. The filtrate was the methyl ester(MS, M⁺+H=300) and the solids were the desired acid, namely,3-nitro-5-(oxazolo[4,5-b]pyridin-2-yl)benzoic acid (MS, M⁺+H=286).

Preparation of3-amino-N-(2-(dimethylamino)ethyl)-5-(oxazolo[4,5-b]pyridin-2-yl)benzamide

3-Nitro-5-(oxazolo[4,5-b]pyridin-2-yl)benzoic acid (250 mg, 0.877 mmol)was dissolved in 5 mL of DMF along with 1 eq ofN,N-dimethylethylenediamine, 500 mg of HATU (1.5 eq) and 0.3 mL of DIEA(2 eq). The reaction mixture was stirred at room temperature for 18hours. It was then diluted with 50 mL of EtOAc and washed with water.The organic layer was dried (Na₂SO₄) and concentrated to afford thenitro amide intermediate. This was dissolved in 50 mL of MeOH and 5 mLof water along with 200 mg of sodium hydrosulfide hydrate (4 eq). Thereaction mixture was stirred under reflux for 1 hour. It was thenconcentrated to dryness and mixed with 50 mL of 1:1 CH₂Cl₂/MeOH andfiltered. The filtrate was concentrated to afford essentiallyquantitative yield of3-amino-N-(2-(dimethylamino)ethyl)-5-(oxazolo[4,5-b]pyridin-2-yl)benzamide(MS, M⁺+H=326).

Preparation of tert-butyl4-(3-amino-5-(oxazolo[4,5-b]pyridin-2-yl)benzoyl)piperazine-1-carboxylate

3-Nitro-5-(oxazolo[4,5-b]pyridin-2-yl)benzoic acid (250 mg, 0.877 mmol)was dissolved in 5 mL of DMF along with 1 eq of Boc-piperazine (163 mg),500 mg of HATU (1.5 eq) and 0.3 mL of DIEA (2 eq). The reaction mixturewas stirred at room temperature for 18 hours. It was then diluted with50 mL of EtOAc and washed with water. The organic layer was dried(Na₂SO₄) and concentrated to afford the nitro amide intermediate. Thiswas dissolved in 50 mL of MeOH and 5 mL of water along with 200 mg ofsodium hydrosulfide hydrate (4 eq). The reaction mixture was stirredunder reflux for 1 hour. The reaction mixture was concentrated and theaqueous layer was extracted with CH₂Cl₂. The combined organic layerswere dried (Na₂SO₄) and concentrated to afford 300 mg of tert-butyl4-(3-amino-5-(oxazolo[4,5-b]pyridin-2-yl)benzoyl)piperazine-1-carboxylate.(MS, M⁺+H=424).

Preparation of Compound 164

Tert-Butyl4-(3-amino-5-(oxazolo[4,5-b]pyridin-2-yl)benzoyl)piperazine-1-carboxylate(100 mg, 0.236 mmol) was dissolved in 1 mL of pyridine along with 47 mgof 3,4-dimethoxybenzoyl chloride (1 eq). The reaction mixture was heatedin a Biotage microwave reactor for 10 min. It was then cooled to roomtemperature and concentrated. The resulting residue was purified bychromatography using a 9:1 mixture of CH₂Cl₂ to MeOH to afford 120 mg ofthe Boc-protected diamide derivative. This was treated with 2 mL of 25%TFA in CH₂Cl₂ and allowed to stand at room temperature for 1 hour. Itwas then concentrated and triturated with Et₂O to afford3,4-dimethoxy-N-(3-(oxazolo[4,5-b]pyridin-2-yl)-5-(piperazine1-carbonyl)phenyl)benzamide as the TFA salt (MS, M⁺+H=488).

Preparation of (3-nitro-5-oxazolo[4,5-b]pyridin-2-yl-phenyl)-methanol

In a typical run, 3-nitro-5-(oxazolo[4,5-b]pyridin-2-yl)benzoic acid(770 mg, 2.70 mmol) was suspended in 50 mL of anhydrous THF and cooledin an ice bath along with NMM (0.3 mL, 2.70 mmol). Isobutylchloroformate (0.35 mL, 2.70 mmol) was added and the reaction mixturewas stirred at 0° C. for 1 hour. A solution of NaBH₄ (102 mg, 2.70 mmol)in 6 mL of water was then added at 0° C. and the reaction mixture wasstirred at the same temperature for 45 min. It was then concentrated andthe resulting residue was diluted with 10 mL of water. The aqueous layerwas extracted with CH₂Cl₂. The organic layer was dried (Na₂SO₄) andconcentrated to afford 500 mg of(3-nitro-5-oxazolo[4,5-b]pyridin-2-yl-phenyl)-methanol (68% yield, >95%pure by LC/MS).

Preparation of4-(3-amino-5-oxazolo[4,5-b]pyridin-2-yl-benzyl)-piperazine-1-carboxylicacid tert-butyl ester

In a typical run, 250 mg of(3-nitro-5-oxazolo[4,5-b]pyridin-2-yl-phenyl)-methanol (0.923 mmol) wasdissolved in 25 mL of CH₂Cl₂ along with 1 eq of Et₃N (130 μL).Methanesulfonyl chloride (1 eq, 70 μL) was added and the reactionmixture was warmed to room temperature and stirred for 15 min. It wasthen quenched with brine and extracted with CH₂Cl₂. The combined organiclayers were dried (Na₂SO₄) and concentrated to afford the mesylateintermediate. This material was mixed with 4 mL of CH₃CN along with 130μL of Et₃N and 172 mg of Boc-piperazine (0.923 mmol) and stirred at roomtemperature for 1 day. The reaction mixture was concentrated and theresulting residue was partitioned between CH₂Cl₂ and water. The organiclayer was dried (Na₂SO₄) and concentrated to afford essentiallyquantitative yield of4-(3-nitro-5-oxazolo[4,5-b]pyridin-2-yl-benzyl)-piperazine-1-carboxylicacid tert-butyl ester. This material was mixed with 6 mL of MeOH and 1mL of water along with 200 mg of sodium hydrosulfide hydrate. Theresulting reaction mixture was stirred under reflux for 1 hour. It wasthen cooled to room temperature and concentrated. The resulting residuewas diluted with 2 mL of water and extracted with CH₂Cl₂. The combinedorganic layers were dried (Na₂SO₄) and concentrated to afford 280 mg of4-(3-amino-5-oxazolo[4,5-b]pyridin-2-yl-benzyl)-piperazine-1-carboxylicacid tert-butyl ester.

Preparation of Compound 159

4-(3-Amino-5-oxazolo[4,5-b]pyridin-2-yl-benzyl)-piperazine-1-carboxylicacid tert-butyl ester (0.2 mmol) was mixed with 1 mL of pyridine alongwith 1 eq (40 mg) of 3,4-dimethoxybenzoyl chloride. The reaction mixturewas reacted in a Biotage microwave reactor at 160° C. for 10 min. It wasthen cooled to room temperature and concentrated. The resulting crudeproduct was purified by chromatography (Isco, gradient elution, CH₂Cl₂to 95% CH₂Cl₂, 4% MeOH and 1% Et₃N). The purified product was thentreated with 2 mL of 25% TFA in CH₂Cl₂ for 2 hours. It was thenconcentrated and the resulting residue was triturated with Et₂O toafford the desired product as the TFA salt (MS, M⁺+H=474).

Preparation of Compound 160 and Compound 161:

The same procedure used in the preparation of Compound 159 was employedusing the appropriate acid chloride.

Preparation of3-dimethylaminomethyl-5-oxazolo[4,5-b]pyridin-2-yl-phenylamine

In a typical run, 250 mg of(3-nitro-5-oxazolo[4,5-b]pyridin-2-yl-phenyl)-methanol (0.923 mmol) wasdissolved in 25 mL of CH₂Cl₂ along with 1 eq of Et₃N (130 μL).Methanesulfonyl chloride (1 eq, 70 μL) was added and the reactionmixture was warmed to room temperature and stirred for 15 min. It wasthen quenched with brine and extracted with CH₂Cl₂. The combined organiclayers were dried (Na₂SO₄) and concentrated to afford the mesylateintermediate. This material was dissolved in 5 mL of CH₃CN along with 2mL of 2 N-dimethylamine in THF. The reaction mixture was stirred at roomtemperature for 2 hours. It was then concentrated and the resultingresidue was partitioned between CH₂Cl₂ and water. The organic layer wasseparated, dried (Na₂SO₄) and concentrated to afford the crude nitroderivative. This material was mixed with 6 mL of MeOH and 1 mL of wateralong with 200 mg of sodium hydrosulfide hydrate and stirred underreflux for 1 hour. The reaction mixture was cooled to room temperatureand diluted with 100 mL of absolute EtOH and concentrated to dryness.The resulting residue was mixed with 10 mL of 9:1 CH₂Cl₂/MeOH andfiltered. The filtrate was concentrated to afford 220 mg of3-dimethylaminomethyl-5-oxazolo[4,5-b]pyridin-2-yl-phenylamine.

Preparation of Compound 156

3-Dimethylaminomethyl-5-oxazolo[4,5-b]pyridin-2-yl-phenylamine (0.2mmol) was mixed with 1 mL of pyridine along with 40 mg of3,4-dimethoxybenzoyl chloride (0.2 mmol). The reaction mixture wasreacted in a Biotage microwave reactor at 160° C. for 10 min. It wasthen cooled to room temperature and concentrated. The resulting residuewas purified by chromatography (Isco, gradient elution, CH₂Cl₂ to 95%CH₂Cl₂, 4% MeOH and 1% Et₃N) to afford the desired product (MS,M⁺+H=433).

Preparation of Compound 157 and Compound 158:

The same procedure used in the preparation of Compound 156 was employedusing the appropriate acid chloride.

Preparation of (3-nitro-5-thiazolo[5,4-c]pyridin-2-yl-phenyl)-methanol

3-Nitro-5-thiazolo[5,4-c]pyridin-2-yl-benzoic acid (880 mg, 2.92 mmol)was suspended in 50 mL of anhydrous THF along with NMM (0.32 mL, 2.92mmol). The reaction mixture was cooled in an ice bath and isobutylchloroformate (0.38 mL, 2.92 mmol) was added. The reaction mixture wasstirred at 0° C. for 40 min. NaBH₄ (110 mg, 2.92 mmol) was then added asa solution in 5 mL of water. The reaction mixture was stirred at 0° C.for 30 min and then warmed to room temperature. It was concentrated andthen extracted with CH₂Cl₂. The combined organic layers were dried(Na₂SO₄) and concentrated to afford the crude product. Purification bychromatography (Isco, gradient elution, CH₂Cl₂ to 9:1 CH₂Cl₂/MeOH)afford 150 mg of(3-nitro-5-thiazolo[5,4-c]pyridin-2-yl-phenyl)-methanol.

Preparation of3-dimethylaminomethyl-5-thiazolo[5,4-c]pyridin-2-yl-phenylamine

(3-Nitro-5-thiazolo[5,4-c]pyridin-2-yl-phenyl)-methanol (120 mg, 0.418mmol) was suspended in 20 mL of CH₂Cl₂ along with Et₃N (87 μL, 1.5 eq)cooled in an ice bath. Methanesulfonyl chloride (32 μL, 0.418 mmol) wasadded and the reaction mixture was slowly warmed to room temperature.The reaction mixture was partitioned between CH₂Cl₂ and brine. Theorganic layer was separated, dried (Na₂SO₄) and concentrated to affordthe crude mesylate intermediate. This material was dissolved in 2 mL ofCH₃CN along with 2 mL of 2 N dimethylamine in THF. The resultingreaction mixture was stirred at room temperature for 2 hours. It wasthen concentrated. The resulting residue was mixed with 6 mL of MeOH and2 mL of water containing 200 mg of sodium hydrosulfide hydrate. Thereaction mixture was stirred under reflux for 3 hours. It was thencooled to room temperature, diluted with 100 mL of absolute EtOH, andconcentrated to dryness. The resulting residue was mixed with 10 mL of9:1 CH₂Cl₂/MeOH and filtered. The filtrate was concentrated to affordessentially quantitative yield of3-dimethylaminomethyl-5-thiazolo[5,4-c]pyridin-2-yl-phenylamine.

Preparation of Compound 174

3-Dimethylaminomethyl-5-thiazolo[5,4-c]pyridin-2-yl-phenylamine (0.2mmol) was mixed with 1 mL of pyridine along with 47 mg of3,4,5-trimethoxybenzoyl chloride (0.2 mmol). The reaction mixture wasreacted in a Biotage microwave reactor at 160° C. for 10 min. It wasthen cooled to room temperature and concentrated. The resulting residuewas purified by chromatography (Isco, gradient elution, CH₂Cl₂ to 95%CH₂Cl₂, 4% MeOH and 1% Et₃N) to afford 30 mg of the desired product (MS,M⁺+H=479).

Preparation of 3-(oxazolo[4,5-b]pyridin-2-yl)benzoic acid

2-Amino-3-hydroxypyridine (1 g, 9.16 mmol) was dissolved in 25 mL of DMFalong with 1.65 g of mono-methyl isophthalate (9.16 mmol), 5.2 g of HATU(1.5 eq) and 3.2 mL of DIEA. The reaction mixture was stirred at roomtemperature for 18 hrs. It was then diluted with 150 mL of ethyl acetateand washed with water (5×20 mL). The organic layer was dried (Na₂SO₄)and concentrated to afford 3.20 g of the intermediate amide. Thismaterial was mixed with 10 mL of PPA and stirred at 160 degree for 4hrs. It was then cooled to room temperature and carefully poured into150 mL of water. The pH was brought to about 5 with solid NaOH. Theresulting precipitate was collected by filtration and dried to afford380 mg of the desired acid, namely,3-(oxazolo[4,5-b]pyridin-2-yl)benzoic acid.

Preparation of Compound 102

For amide formation, 3-(oxazolo[4,5-b]pyridin-2-yl)benzoic acid (30 mg,0.125 mmol) was dissolved in 2 mL of DMF along with3,4,5-trimethoxyaniline (23 mg, 1 eq) and 71 mg of HATU (1.5 eq). After45 microliters of DIEA (2 eq) were added, the reaction mixture wasstirred at room temperature for 18 hrs. It was then diluted with ethylacetate and washed with water and concentrated. The resulting residuewas purified by chromatography using a 9:1 mixture of CH₂Cl₂ to MeOH toafford3-(oxazolo[4,5-b]pyridin-2-yl)-N-(3,4,5-trimethoxyphenyl)benzamide. (MS,M⁺+H=406).

Preparation of Compound 100, Compound 101 and Compound 103:

The same procedure used in the preparation of Compound 102 was employedusing the appropriate amine.

Preparation of 5-oxazolo[4,5-b]pyridin-2-yl-pyridin-3-ylamine

In a typical run, 790 mg of 2-amino-3-hydroxypyridine (7.24 mmol) and1.00 g of 5-aminonicotinic acid (7.24 mmol) were mixed with 10 mL of PPAand stirred at 200° C. for 6 hours. The reaction mixture was cooled toabout 100° C. and carefully poured into 100 mL of water. The pH wasbrought to 6 with solid NaOH and the solids were collected byfiltration. After drying under high vacuum, a total of 180 mg of5-oxazolo[4,5-b]pyridin-2-yl-pyridin-3-ylamine.

Preparation of Compound 73

5-Oxazolo[4,5-b]pyridin-2-yl-pyridin-3-ylamine (25 mg, 0.118 mmol) wasmixed with 1 mL of pyridine along with 24 mg of 2,4-dimethoxybenzoylchloride (0.118 mmol). The reaction mixture was reacted in a Biotagemicrowave reactor at 160° C. for 10 min. It was then cooled to roomtemperature and concentrated. The resulting crude product was purifiedby chromatography (Isco, gradient elution, CH₂Cl₂ to 9:1 CH₂Cl₂/MeOH) toafford 11 mg of the product (MS, M⁺+H=377).

Preparation of Compounds 66, 67 and 68:

Essentially the same procedure as detailed for the preparation ofCompound 73 was used employing the appropriate acid chloride.

Preparation of 2-imidazo[2,1-b]thiazol-6-yl-phenylamine

In a typical run, 123 mg of 2-aminothiazole (1.23 mmol) and2-bromo-2′-nitroacetophenone (300 mg, 1.23 mmol) was mixed with 15 mL ofmethyl ethyl ketone and stirred under reflux for 18 hours. It was thencooled to room temperature and filtered. The filtrate was concentrated.The resulting solids were mixed with 20 mL of EtOH and 5 drops ofconcentrated HBr were added. The reaction mixture was stirred underreflux for 6 hours. Everything dissolved at this point and LC/MS showedformation of desired nitro intermediate (MS, M⁺+H=246). The reactionmixture was concentrated and mixed with 20 mL of dilute aq. NaHCO₃. Theresulting solids were collected by filtration and dried to afford 300 mgof the nitro intermediate. This material was mixed with 15 mL of MeOHand 3 mL of water along with 6 eq of sodium hydrosulfide hydrate. Thereaction mixture was stirred under reflux for 8 hours. It was thencooled to room temperature and concentrated. The aqueous layer wasextracted with CH₂Cl₂. The combined organic layers were dried (Na₂SO₄)and concentrated to afford 260 mg of2-imidazo[2,1-b]thiazol-6-yl-phenylamine.

Preparation of Compound 203

2-Imidazo[2,1-b]thiazol-6-yl-phenylamine (64 mg, 0.30 mmol) was mixedwith 1 mL of pyridine along with 60 mg of 3,4-dimethoxybenzoyl chloride(0.30 mmol). The reaction mixture was reacted in the Biotage microwavereactor at 160° C. for 10 min. The reaction mixture was cooled to roomtemperature and concentrated. The resulting residue was purified bychromatography (Isco, gradient elution, CH₂Cl₂ to 9:1 CH₂Cl₂/MeOH) toafford the desired product (MS, M⁺+H=380).

Preparation of Compound 204:

The same procedure used in the preparation of Compound 203 was employedusing the appropriate acid chloride.

Preparation of Compounds 707, 739 and 740

The same procedure used in the preparation of Compound 203 was employedexcept with 2-amino-4-methylthiazole at the beginning of the syntheticsequence, and the appropriate acid chloride at the last amide formationstep.

Preparation of 6-(2-Nitro-phenyl)-imidazo[2,1-b]thiazole-3-carboxylicacid ethyl ester

In a typical run, 2.1 g of ethyl 2-aminothiazole-4-carboxylate(Combi-Blocks, 0.0123 mol) was mixed with 25 mL of methyl ethyl ketonealong with 2-bromo-2′-nitroacetophenone (3.0 g, 0.0123 mol). Thereaction mixture was stirred under reflux for 18 hours. It was thencooled to room temperature and filtered to remove some of the solids.The filtrate was concentrated to afford 3.10 g of6-(2-nitro-phenyl)-imidazo[2,1-b]thiazole-3-carboxylic acid ethyl ester(MS, M⁺+H=318).

Preparation of [6-(2-nitro-phenyl)-imidazo[2,1-b]thiazol-3-yl]-methanol

6-(2-Nitro-phenyl)-imidazo[2,1-b]thiazole-3-carboxylic acid ethyl ester(14.50 g, 0.0458 mol) was mixed with 100 mL of THF and 100 mL of watercontaining 7.3 g of NaOH (4 eq). The reaction mixture was stirred atroom temperature for 18 hours. It was then concentrated. The aqueouslayer was washed once with CH₂Cl₂ and then acidified with 6 N HCl. Thesolids were collected by filtration and dried to afford 7.4 g of theacid intermediate. This material (7.4 g, 0.0256 mol) was mixed with 200mL of anhydrous THF along with NMM (2.8 mL, 0.0256 mol) and cooled to 0°C. Isobutyl chloroformate (3.35 mL, 0.0256 mol) was added and thereaction mixture was stirred in the ice bath for 3 hours. NaBH₄ (0.97 g,0.0256 mol) was added as a solution in 30 mL of water. The reactionmixture was stirred at 0° C. for 45 min. It was then warmed to roomtemperature and concentrated. The aqueous layer was extracted withCH₂Cl₂. The combined organic layers were dried (Na₂SO₄) and concentratedto afford the crude product. Purification by chromatography (Isco, usinga mixture of pentane/EtOAc) afforded 5.20 g of[6-(2-nitro-phenyl)-imidazo[2,1-b]thiazol-3-yl]-methanol (74% yield).

Preparation of4-[6-(2-amino-phenyl)-imidazo[2,1-b]thiazol-3-ylmethyl]-piperazine-1-carboxylicacid tert-butyl ester

[6-(2-Nitro-phenyl)-imidazo[2,1-b]thiazol-3-yl]-methanol (1.0 g, 3.64mmol) was dissolved in 100 mL of CH₂Cl₂ along with 1 eq of Et₃N (0.51mL). Methanesulfonyl chloride (1 eq, 0.28 mL) was added and the reactionmixture was warmed to room temperature and stirred for 15 min. It wasthen quenched with brine and extracted with CH₂Cl₂. The combined organiclayers were dried (Na₂SO₄) and concentrated to afford the mesylateintermediate. This material was mixed with 4 mL of CH₃CN along with 0.51mL of Et₃N and 680 mg of Boc-piperazine (3.64 mmol) and stirred at roomtemperature for 1 day. The reaction mixture was concentrated and theresulting residue was partitioned between CH₂Cl₂ and water. The organiclayer was dried (Na₂SO₄) and concentrated to afford essentiallyquantitative yield of the product. This material was mixed with 6 mL ofMeOH and 1 mL of water along with 200 mg of sodium hydrosulfide hydrate.The resulting reaction mixture was stirred under reflux for 24 hours. Itwas then cooled to room temperature and concentrated. The resultingresidue was diluted with 2 mL of water and extracted with CH₂Cl₂. Thecombined organic layers were dried (Na₂SO₄) and concentrated to afford0.90 g of4-[6-(2-amino-phenyl)-imidazo[2,1-b]thiazol-3-ylmethyl]-piperazine-1-carboxylicacid tert-butyl ester.

Preparation of Compound 207

4-[6-(2-Amino-phenyl)-imidazo[2,1-b]thiazol-3-ylmethyl]-piperazine-1-carboxylicacid tert-butyl ester (0.3 mmol) was mixed with 1 mL of pyridine alongwith 1 eq (60 mg) of 3,4-dimethoxybenzoyl chloride. The reaction mixturewas reacted in a Biotage microwave reactor at 160° C. for 10 min. It wasthen cooled to room temperature and concentrated. The resulting crudeproduct was purified by chromatography (Isco, gradient elution, CH₂Cl₂to 95% CH₂Cl₂, 4% MeOH and 1% Et₃N). The purified product was thentreated with 2 mL of 25% TFA in CH₂Cl₂ for 2 hours. It was thenconcentrated and the resulting residue was triturated with Et₂O toafford the desired product as the TFA salt (MS, M⁺+H=478).

Preparation of Compounds 208, 326, 327, 328, 329, 330, 337, 338, 440,441, 442, 443, 444, 445, 446, 447, 448, 510, 511, 512, 543, 544, 708,709, 710, 733, 735, 736, 737, 738, 743 and 744

The same procedure used in the preparation of Compound 207 was employedusing the appropriate acid chloride or sulfonyl chloride. Compounds 623,624, 625, 644, 645, 692, 695, 697 and 698 were prepared according to theprocedure used for the preparation of Compound 207, using theappropriate acid chloride. The acid chlorides were either commerciallyavailable or made from the carboxylic acids as follows: The carboxylicacid (1.0 mmol), thionyl chloride (2.0 mmol), and a catalytic amount ofN,N-dimethylformamide (DMF) (2 drops) was refluxed in toluene (2 mL) for1 hour. The reaction was cooled to room temperature and concentrated invacuo to afford the desired acid chlorides.

Preparation of2-(3-Dimethylaminomethyl-imidazo[2,1-b]thiazol-6-yl)-phenylamine

[6-(2-Nitro-phenyl)-imidazo[2,1-b]thiazol-3-yl]-methanol (435 mg, 1.58mmol) was dissolved in 25 mL of CH₂Cl₂ along with 1 eq of Et₃N (0.330mL). Methanesulfonyl chloride (1 eq, 0.12 mL) was added and the reactionmixture was warmed to room temperature and stirred for 15 min. It wasthen quenched with brine and extracted with CH₂Cl₂. The combined organiclayers were dried (Na₂SO₄) and concentrated to afford the mesylateintermediate. This material was mixed with 4 mL of THF along with 4 mLof a 2 N dimethylamine solution in THF and stirred at room temperaturefor 3 hrs. The reaction mixture was concentrated and the resultingresidue was partitioned between CH₂Cl₂ and water. The organic layer wasdried (Na₂SO₄) and concentrated to afford essentially quantitative yieldof the product. This material was mixed with 6 mL of MeOH and 1 mL ofwater along with 200 mg of sodium hydrosulfide hydrate. The resultingreaction mixture was stirred under reflux for 6 hours. It was thencooled to room temperature, diluted with 100 mL of absolute EtOH andconcentrated. The resulting residue was mixed with 20 mL of 9:1CH₂Cl₂/MeOH and filtered. The filtrate was concentrated to afford2-(3-dimethylaminomethyl-imidazo[2,1-b]thiazol-6-yl)-phenylamine.

Preparation of Compound 205

2-(3-Dimethylaminomethyl-imidazo[2,1-b]thiazol-6-yl)-phenylamine (0.3mmol) was mixed with 1 mL of pyridine along with 1 eq (60 mg) of3,4-dimethoxybenzoyl chloride. The reaction mixture was reacted in aBiotage microwave reactor at 160° C. for 10 min. It was then cooled toroom temperature and concentrated. The resulting crude product waspurified by chromatography (Isco, gradient elution, CH₂Cl₂ to 95%CH₂Cl₂, 4% MeOH and 1% Et₃N) to afford the desired product as a lightyellow solid (MS, M⁺+H=437).

Preparation of Compound 206

The same procedure used in the preparation of Compound 205 was employedusing the appropriate acid chloride.

Preparation of2-(3-morpholin-4-ylmethyl-imidazo[2,1-b]thiazol-6-yl)-phenylamine

[6-(2-Nitro-phenyl)-imidazo[2,1-b]thiazol-3-yl]-methanol (435 mg, 1.58mmol) was dissolved in 25 mL of CH₂Cl₂ along with 1 eq of Et₃N (0.330mL). Methanesulfonyl chloride (1 eq, 0.12 mL) was added and the reactionmixture was warmed to room temperature and stirred for 15 min. It wasthen quenched with brine and extracted with CH₂Cl₂. The combined organiclayers were dried (Na₂SO₄) and concentrated to afford the mesylateintermediate. This material was mixed with 6 mL of CH₃CN along with 0.33mL of Et₃N and 0.14 mL of morpholine. The reaction mixture was stirredat room temperature for 18 hours. The next day, it was concentrated andthe resulting residue was partitioned between CH₂Cl₂ and water. Theorganic layer was dried (Na₂SO₄) and concentrated to afford essentiallyquantitative yield of the product. This material was mixed with 6 mL ofMeOH and 1 mL of water along with 200 mg of sodium hydrosulfide hydrate.The resulting reaction mixture was stirred under reflux for 6 hours. Itwas then cooled to room temperature, diluted with 100 mL of absoluteEtOH and concentrated. The resulting residue was mixed with 20 mL of 9:1CH₂Cl₂/MeOH and filtered. The filtrate was concentrated to afford2-(3-morpholin-4-ylmethyl-imidazo[2,1-b]thiazol-6-yl)-phenylamine.

Preparation of Compound 209

2-(3-Morpholin-4-ylmethyl-imidazo[2,1-b]thiazol-6-yl)-phenylamine (0.3mmol) was mixed with 1 mL of pyridine along with 1 eq (60 mg) of3,4-dimethoxybenzoyl chloride. The reaction mixture was reacted in aBiotage microwave reactor at 160° C. for 10 min. It was then cooled toroom temperature and concentrated. The resulting crude product waspurified by chromatography (Isco, gradient elution, CH₂Cl₂ to 95%CH₂Cl₂, 4% MeOH and 1% Et₃N) to afford the desired product as a lightyellow solid (MS, M⁺+H=479).

Preparation of Compound 210

The same procedure used in the preparation of Compound 209 was employedusing the appropriate acid chloride.

Preparation of 6-(2-nitro-phenyl)-imidazo[2,1-b]thiazole-2-carboxylicacid ethyl ester

In a typical run, 1.0 g of ethyl 2-aminothiazole-5-carboxylate(Astatech, 5.81 mmol) was mixed with 50 mL of acetone along with 1.42 gof 2-bromo-2′-nitroacetophenone and stirred under reflux for 18 hours.It was then filtered. The filtrate was concentrated to afford theintermediate amide (MS, M⁺+H=336). This material was mixed with 20 mL ofEtOH along with 6 drops of concentrated HBr and stirred under reflux for4 hours. The reaction mixture was cooled to room temperature andconcentrated. The resulting residue was diluted with dilute aqueousNaHCO₃. The solids were collected by filtration and dried to afford6-(2-nitro-phenyl)-imidazo[2,1-b]thiazole-2-carboxylic acid ethyl ester(MS, M⁺+H=318).

Preparation of [6-(2-nitro-phenyl)-imidazo[2,1-b]thiazol-2-yl]-methanol

6-(2-Nitro-phenyl)-imidazo[2,1-b]thiazole-2-carboxylic acid ethyl ester(660 mg, 2.08 mmol) was dissolved in 12 mL of THF and NaOH (4 eq) wasadded as a solution in 10 mL of water. The reaction mixture was stirredat 50° C. for 12 hours. It was then cooled to room temperature andconcentrated. The aqueous layer was acidified to pH 5 with 6 N HCl. Thesolids were collected by filtration and dried to afford essentiallyquantitative yield of the acid. This material (2.08 mmol) was mixed with20 mL of anhydrous THF along with NMM (0.23 mL, 2.08 mmol) and cooled inan ice bath. Isobutyl chloroformate (0.27 mL, 2.08 mmol) was added andthe reaction mixture was stirred at 0° C. for 30 min. NaBH₄ (80 mg, 2.08mmol) was added as a solution in 5 mL of water. The reaction mixture wasstirred at 0° C. for 30 min and then concentrated. The aqueous layer wasextracted with CH₂Cl₂. The combined organic layers were dried (Na₂SO₄)and concentrated. Purification by chromatography (Isco, gradient elutionusing a mixture of CH₂ Cl₂ and MeOH) afforded 190 mg of[6-(2-nitro-phenyl)-imidazo[2,1-b]thiazol-2-yl]-methanol.

Preparation of2-(2-Dimethylaminomethyl-imidazo[2,1-b]thiazol-6-yl)-phenylamine

Essentially the same procedure used during the preparation of2-(3-dimethylaminomethyl-imidazo[2,1-b]thiazol-6-yl)-phenylamine wasemployed, except that[6-(2-nitro-phenyl)-imidazo[2,1-b]thiazol-2-yl]-methanol was used as thestarting material.

Preparation of Compound 178

2-(2-Dimethylaminomethyl-imidazo[2,1-b]thiazol-6-yl)-phenylamine (0.3mmol) was mixed with 1 mL of pyridine along with 1 eq (60 mg) of3,4-dimethoxybenzoyl chloride. The reaction mixture was reacted in aBiotage microwave reactor at 160° C. for 10 min. It was then cooled toroom temperature and concentrated. The resulting crude product waspurified by chromatography (Isco, gradient elution, CH₂Cl₂ to 95%CH₂Cl₂, 4% MeOH and 1% Et₃N) to afford the desired product as a lightyellow solid (MS, M⁺+H=437).

Preparation of Compound 179

The same procedure used in the preparation of Compound 178 was employedusing the appropriate acid chloride.

Preparation of4-[6-(2-Amino-phenyl)-imidazo[2,1-b]thiazol-2-ylmethyl]-piperazine-1-carboxylicacid tert-butyl ester

Essentially the same procedure used during the preparation of4-[6-(2-amino-phenyl)-imidazo[2,1-b]thiazol-3-ylmethyl]-piperazine-1-carboxylicacid tert-butyl ester was employed, except that[6-(2-nitro-phenyl)-imidazo[2,1-b]thiazol-2-yl]-methanol was used as thestarting material.

Preparation of Compound 270

4-[6-(2-Amino-phenyl)-imidazo[2,1-b]thiazol-2-ylmethyl]-piperazine-1-carboxylicacid tert-butyl ester (0.2 mmol) was mixed with 1 mL of pyridine alongwith 1 eq (40 mg) of 3,4-dimethoxybenzoyl chloride. The reaction mixturewas reacted in a Biotage microwave reactor at 160° C. for 10 min. It wasthen cooled to room temperature and concentrated. The resulting crudeproduct was purified by chromatography (Isco, gradient elution, CH₂Cl₂to 95% CH₂Cl₂, 4% MeOH and 1% Et₃N). The purified product was thentreated with 2 mL of 25% TFA in CH₂Cl₂ for 2 hours. It was thenconcentrated and the resulting residue was triturated with Et₂O toafford the desired product as the TFA salt (MS, M⁺+H=478).

Preparation of Compound 271 and Compound 513

The same procedure used in the preparation of Compound 270 was employedusing the appropriate acid chloride.

Preparation of 3-(2-nitro-phenyl)-imidazo[2,1-b]thiazole-6-carboxylicacid ethyl ester

4-(2-Nitro-phenyl)-thiazol-2-ylamine was prepared as follows:2-Bromo-2′-nitroacetophenone (1.75 g, 7.2 mmol) was mixed with 50 mL ofabsolute EtOH along with thiourea (1.09 g, 14.4 mmol) and stirred underreflux for 2 hours. The reaction mixture was cooled to room temperatureand concentrated under reduced pressure. The resulting residue wasbasified with 20 mL of 1 N aqueous NaOH and extracted with CH₂Cl₂. Thecombined organic layers were dried (Na₂SO₄) and concentrated to affordessentially quantitative yield of 4-(2-nitro-phenyl)-thiazol-2-ylamine.

4-(2-Nitro-phenyl)-thiazol-2-ylamine (1.60 g, 7.2 mmol) was mixed with50 mL of methyl ethyl ketone along 0.90 mL of ethyl bromopyruvate (7.2mmol) and stirred under reflux for 24 hours. Another equivalent of ethylbromopyruvate (0.90 mL, 7.2 mmol) was added and the reaction mixture wasstirred under reflux for another 8 hours. The reaction mixture wascooled to room temperature and concentrated under reduced pressure. Theresulting residue was purified by chromatography (Isco, gradientelution, CH₂Cl₂ to 95% CH₂Cl₂, 5% MeOH) to afford 1.2 g of3-(2-nitro-phenyl)-imidazo[2,1-b]thiazole-6-carboxylic acid ethyl ester(52% yield).

Preparation of [3-(2-nitro-phenyl)-imidazo[2,1-b]thiazol-6-yl]-methanol

3-(2-Nitro-phenyl)-imidazo[2,1-b]thiazole-6-carboxylic acid ethyl ester(1.2 g, 3.78 mmol) was mixed with 50 mL of THF and 50 mL of watercontaining 600 mg of NaOH (4 eq). The reaction mixture was stirred at50° C. for 8 hours. It was then cooled to room temperature andconcentrated under reduced pressure. The aqueous layer was acidified topH 6 with 6 N HCl. The resulting solids were collected by filtration anddried to afford 465 mg of the intermediate acid. This acid intermediate(465 mg, 1.61 mmol) was mixed with 50 mL of anhydrous THF along with NMM(0.18 mL, 1.61 mmol). The reaction mixture was cooled in an ice bath andisobutyl chloroformate (0.21 mL, 1.61 mmol) was added. After 30 minutesat 0° C., a solution of NaBH₄ (240 mg) in 2 mL of water was added. Theresulting reaction mixture was stirred at 0° C. for 30 min and thenconcentrated. The aqueous layer was extracted with CH₂Cl₂. The combinedorganic layers were dried (Na₂SO₄) and concentrated to afford the crudealcohol. Purification by chromatography (Isco, gradient elution, CH₂Cl₂to 95% CH₂Cl₂, 5% MeOH) afforded 200 mg of[3-(2-nitro-phenyl)-imidazo[2,1-b]thiazol-6-yl]-methanol (45% yield).

Preparation of4-[3-(2-nitro-phenyl)-imidazo[2,1-b]thiazol-6-ylmethyl]-piperazine-1-carboxylicacid tert-butyl ester

[3-(2-Nitro-phenyl)-imidazo[2,1-b]thiazol-6-yl]-methanol (200 mg, 0.727mmol) was mixed with 50 mL of CH₂Cl₂ along with Et₃N (0.10 mL, 0.727mmol) and cooled in an ice bath. Methanesulfonyl chloride (56 μL, 0.727mmol) was added and the reaction mixture was warmed to room temperatureand stirred for 30 min. The reaction mixture was quenched with brine andextracted with CH₂Cl₂. The combined organic layers were dried (Na₂SO₄)and concentrated to afford essentially quantitative yield of themesylate intermediate. This material was dissolved in 10 mL ofacetonitrile along with triethylamine (0.10 mL, 0.727 mmol) andBoc-piperazine. The reaction mixture was stirred at room temperature for18 hours. It was then concentrated. The resulting residue waspartitioned between water and CH₂Cl₂. The organic layer was separated,dried (Na₂SO₄) and concentrated to afford essentially quantitative yieldof4-[3-(2-nitro-phenyl)-imidazo[2,1-b]thiazol-6-ylmethyl]-piperazine-1-carboxylicacid tert-butyl ester.

Preparation of4-[3-(2-amino-phenyl)-imidazo[2,1-b]thiazol-6-ylmethyl]-piperazine-1-carboxylicacid tert-butyl ester

4-[3-(2-Nitro-phenyl)-imidazo[2,1-b]thiazol-6-ylmethyl]-piperazine-1-carboxylicacid tert-butyl ester (320 mg, 0.73 mmol) was mixed with 20 mL of MeOHand 5 mL of water containing sodium hydrosulfide hydrate (244 mg, 6 eq).The reaction mixture was stirred under reflux for 4 hours. It was thencooled to room temperature and concentrated. The aqueous layer wasextracted with CH₂Cl₂. The combined organic layers were dried (Na₂SO₄)and concentrated to afford 280 mg of4-[3-(2-amino-phenyl)-imidazo[2,1-b]thiazol-6-ylmethyl]-piperazine-1-carboxylicacid tert-butyl ester (92% yield).

Preparation of Compound 560

4-[3-(2-amino-phenyl)-imidazo[2,1-b]thiazol-6-ylmethyl]-piperazine-1-carboxylicacid tert-butyl ester (45 mg, 0.1 mmol) was mixed with 1 mL of pyridinealong with 1 eq (40 mg) of 2-quinoxaloyl chloride. The reaction mixturewas reacted in a Biotage microwave reactor at 160° C. for 10 min. It wasthen cooled to room temperature and concentrated. The resulting crudeproduct was purified by chromatography (Isco, gradient elution, CH₂Cl₂to 95% CH₂Cl₂, 4% MeOH and 1% Et₃N). The purified product was thentreated with 2 mL of 25% TFA in CH₂Cl₂ for 2 hours. It was thenconcentrated and the resulting residue was triturated with Et₂O toafford the desired product as the TFA salt (MS, M⁺+H=470).

Preparation of Compound 559

The same procedure used in the preparation of Compound 560 was employedusing the appropriate acid chloride.

Preparation of a 1:1 mixture of6-(2-Chloro-pyridin-3-yl)-imidazo[2,1-b]thiazole-3-carboxylic acid ethylester and 6-(2-bromo-pyridin-3-yl)-imidazo[2,1-b]thiazole-3-carboxylicacid ethyl ester

The 1:1 mixture of 2-bromo-1-(2-chloro-pyridin-3-yl)-ethanone and2-bromo-1-(2-bromo-pyridin-3-yl)-ethanone was prepared according to theprocedure outlined in WO 2005/061476. This mixture (5.6 g, approximately0.0240 mol) was mixed with 150 mL of methyl ethyl ketone along with2-amino-thiazole-4-carboxylic acid ethyl ester (4.6 g) and stirred underreflux for 18 hours. The reaction mixture was concentrated. Theresulting residue was mixed with 150 mL of CH₂Cl₂ and filtered. Thefiltered solids were unreacted 2-amino-thiazole-4-carboxylic acid ethylester. The filtrate was concentrated to afford an essentially pure and1:1 mixture of6-(2-Chloro-pyridin-3-yl)-imidazo[2,1-b]thiazole-3-carboxylic acid ethylester and 6-(2-bromo-pyridin-3-yl)-imidazo[2,1-b]thiazole-3-carboxylicacid ethyl ester (3.0 g total).

Preparation of a 1:1 mixture of6-(2-Chloro-pyridin-3-yl)-3-morpholin-4-ylmethyl-imidazo[2,1-b]thiazoleand6-(2-bromo-pyridin-3-yl)-3-morpholin-4-ylmethyl-imidazo[2,1-b]thiazole

The 1:1 mixture of6-(2-Chloro-pyridin-3-yl)-imidazo[2,1-b]thiazole-3-carboxylic acid ethylester and 6-(2-bromo-pyridin-3-yl)-imidazo[2,1-b]thiazole-3-carboxylicacid ethyl ester (3.0 g) was mixed with 100 mL of THF along with 25 mLof water containing 3 g of NaOH. The reaction mixture was stirred at 50°C. for 3 hours. It was then cooled to room temperature and concentrated.The aqueous layer was acidified to pH 5 with 6 N HCl and the resultingmixture was filtered. The solids were collected to afford 2.14 g theintermediate acid.

This 1:1 mixture of the acid (2.14 g) was mixed with 250 mL of anhydrousTHF along with NMM (0.85 mL) and cooled in an ice bath. Isobutylchloroformate (1.0 mL) was added and the reaction mixture was warmed toroom temperature and stirred for 3 hours. The reaction mixture wascooled in an ice bath and NaBH₄ (0.29 g) was added as a solution in 20mL of water. The reaction mixture was stirred for 30 min and then warmedto room temperature. It was concentrated and subsequently extracted withCH₂Cl₂. The combined organic layers were dried (Na₂SO₄) and concentratedto afford 1.5 g of the intermediate alcohol.

This 1:1 mixture of the intermediate alcohol (1.5 g) was mixed with 100mL of CH₂Cl₂ along with Et₃N (0.80 mL) and cooled in an ice bath.Methanesulfonyl chloride (0.44 mL) was added and the reaction mixturewas warmed to room temperature. The reaction mixture was quenched withbrine and the two layers were separated. The organic layer was dried(Na₂SO₄) and concentrated to afford the intermediate mesylate. Thismaterial was immediately mixed with 30 mL of CH₃CN along with 0.80 mL ofEt₃N and 0.5 mL of morpholine. The reaction mixture was stirred at 50°C. for 3 hours. The reaction mixture was concentrated under reducedpressure and the resulting residue was partitioned between CH₂Cl₂ andbrine. The organic layer was separated, dried (Na₂SO₄) and concentratedto afford the crude product. Purification by chromatography (Isco,gradient elution, CH₂Cl₂ to 95% CH₂Cl₂, 4% MeOH and 1% Et₃N) afforded720 mg of a 1:1 mixture of6-(2-Chloro-pyridin-3-yl)-3-morpholin-4-ylmethyl-imidazo[2,1-b]thiazoleand6-(2-bromo-pyridin-3-yl)-3-morpholin-4-ylmethyl-imidazo[2,1-b]thiazole.

Preparation of(4-methoxy-benzyl)-[3-(3-morpholin-4-ylmethyl-imidazo[2,1-b]thiazol-6-yl)-pyridin-2-yl]-amine

The 1:1 mixture of6-(2-Chloro-pyridin-3-yl)-3-morpholin-4-ylmethyl-imidazo[2,1-b]thiazoleand6-(2-bromo-pyridin-3-yl)-3-morpholin-4-ylmethyl-imidazo[2,1-b]thiazole(600 mg) was mixed with 15 mL of toluene along with 0.47 mL of4-methoxybenzylamine and stirred under reflux for 5 days. The reactionmixture was cooled to room temperature and partitioned between CH₂Cl₂and brine. The organic layer was separated, dried (Na₂SO₄) andconcentrated to afford the crude product. Purification by chromatography(Isco, gradient elution, CH₂Cl₂ to 95% CH₂Cl₂, 4% MeOH and 1% Et₃N)afforded 200 mg of(4-methoxy-benzyl)-[3-(3-morpholin-4-ylmethyl-imidazo[2,1-b]thiazol-6-yl)-pyridin-2-yl]-amine.

Preparation of3-(3-morpholin-4-ylmethyl-imidazo[2,1-b]thiazol-6-yl)-pyridin-2-ylamine

(4-Methoxy-benzyl)-[3-(3-morpholin-4-ylmethyl-imidazo[2,1-b]thiazol-6-yl)-pyridin-2-yl]-amine(100 mg, 0.23 mmol) was mixed with 2 mL of CH₂Cl₂ along withtriethylsilane (0.11 mL, 2 eq). Trifluoroacetic acid (1 mL) was addedand the reaction mixture was stirred at room temperature for 18 hours.The following day, the reaction mixture was concentrated. The resultingresidue was triturated with Et₂O to afford essentially quantitativeyield of3-(3-morpholin-4-ylmethyl-imidazo[2,1-b]thiazol-6-yl)-pyridin-2-ylamineas the TFA salt.

Preparation of Compound 621

The TFA salt of3-(3-morpholin-4-ylmethyl-imidazo[2,1-b]thiazol-6-yl)-pyridin-2-ylamine(0.1 mmol) was mixed with 1 mL of pyridine along with 0.1 mmol of2-quinoxaloyl chloride. The reaction mixture was reacted in a microwavereactor at 160° C. for 10 min. It was then cooled to room temperatureand concentrated to afford the crude product. Purification bypreparative HPLC using a mixture of aqueous CH₃CN that has been bufferedwith 0.1% TFA afforded 18 mg of the desired product as the TFA salt (MS,M⁺+H=472).

Preparation of N-(4-hydroxypyridin-3-yl)-3-nitrobenzamide

A suspension of 4-hydroxy-3-nitropyridine (40 g) and 10% Pd/C (4 g) inEtOH (700 mL) and dichloromethane (50 mL) was stirred under H₂ (1 atm)at room temperature for 4 days. TLC indicated completion of thereaction. The reaction mixture was filtered through a Celite pad, andthe filtrate was concentrated in vacuo to give crude3-amino-4-hydroxypyridine as a red foam (32 g, yield: 100%, confirmed byMS), which was used directly for the next step.

A solution of 3-nitrobenzoyl chloride (3.339 g, 18.0 mmol) in pyridine(54.0 mL) was added dropwise to a solution of 3-amino-4-hydroxypyridine(2.376 g, 21.6 mmol) in pyridine (36.0 mL) at 10° C. and the resultantmixture was stirred overnight. Na₂CO₃ (1.145 g, 10.8 mmol) was added andthe mixture was stirred for 1 h. The solid was collected by filtration,washed with 10% HOAc (30 mL×3) and water (30 mL×3), and dried undervacuo to afford N-(4-hydroxypyridin-3-yl)-3-nitrobenzamide as yellowsolid (3.70 g, yield: 66%). ¹H-NMR (400 MHz, DMSO-dB_(6B)) δ: 6.34 (1H,d, J=7.2 Hz), 7.74 (1H, d, J=8.0 Hz), 7.84 (1H, t, J=8.0 Hz), 8.34 (1H,d, J=7.2 Hz), 8.44 (1H, t, J=8.0 Hz), 8.66 (1H, s), 8.73 (1H, s), 9.66(1H, s), 11.65 (1H, br s); MS (ESI) calcd. for C₁₂H₉N₃O₄ (m/z): 259,found: 260 [M+1]⁺.

Preparation of 2-(3-nitrophenyl)oxazolo[4,5-c]pyridine

A solution of N-(4-hydroxypyridin-3-yl)-3-nitrobenzamide (1.554 g, 6mmol) in polyphosphoric acid (12.0 mL) was stirred at 150° C. for 6 h.The reaction mixture was poured into distilled water and sodiumhydroxide was added until pH=5. The precipitate was collected byfiltration, washed with water until neutral and dried in vacuo oven (50°C.) to afford 2-(3-nitrophenyl)oxazolo[4,5-c]pyridine as a yellow solid(1.389 g, yield: 96%). MS (ESI) calcd. for C₁₂H₇NO₃ (m/z): 241, found:242 [M+1]⁺.

Preparation of 3-(oxazolo[4,5-c]pyridin-2-yl)benzenamine

A suspension of 2-(3-nitrophenyl)oxazolo[4,5-c]pyridine (1.50 g, 6.2mmol), iron powder (1.867 g, 31.8 mmol) and NH₄Cl (2.86 g, 53.5 mmol) inCH₃OH/H₂O (4:1, 311.2 mL) was refluxed for 6 h. The mixture was filteredand the filtrate was evaporated in vacuo. The residue was purified bychromatography on silica gel (eluted with petroleumether:EtOAc:Et₃N=160:40:1) to afford3-(oxazolo[4,5-c]pyridin-2-yl)benzenamine as a white solid (1.107 g,yield: 85%). ¹H NMR (400 MHz, DMSO-d₆) δ: 5.54 (2H, s), 6.82 (1H, d,J=8.0 Hz), 7.24 (1H, t, J=8.0 Hz), 7.35 (1H, d, J=8.0 Hz), 7.44 (1H, s),7.86 (1H, d, J=5.6 Hz), 8.56 (1H, d, J=5.6 Hz), 9.07 (1H, s); MS (ESI)calcd. for C₁₂H₉N₃O (m/z): 211, found: 212 [M+1]⁺.

General Procedure for Preparing Compounds 296, 297, 298, 299, 311, 343,344, 345, 346, 347, and 348

The acid chlorides, in turn, were either commercially available orprepared from the corresponding carboxylic acid as follows: 1.0 g of thecarboxylic acid was refluxed in 10 mL of thionyl chloride and 0.1 mL ofDMF for 2 hours. The reaction mixture was cooled to room temperature andconcentrated under reduced pressure to afford the desired acid chloride.A mixture of the 3-(oxazolo[4,5-c]pyridin-2-yl)benzenamine (0.2 mmoleach) and the appropriate acid chloride (0.24 mmol each) in pyridine (2mL) was agitated at room temperature overnight. The reaction mixture wasdiluted with H₂O (5 mL each). The precipitates were collected byfiltration and triturated with MeOH (5 mL) and dried to give librarycompounds, which were analyzed by HPLC & MS. The library compounds werefurther purified by passing through silica gel pad eluted withCH₂Cl₂/EtOAc or petroleum ether/EtOAc.

Preparation of 2-(3-nitrophenyl)thiazolo[4,5-c]pyridine

A mixture of N-(4-hydroxypyridin-3-yl)-3-nitrobenzamide (1.33 g, 5.2mmol) and P₂S₅ (2.40 g, 10.4 mmol) in pyridine (6.0 mL) and p-xylene (24mL) was stirred at 140° C. for 18 h. The solvent was removed under vacuowhile hot and the residue was purified by recrystallization from EtOH toafford 2-(3-nitrophenyl)thiazolo[4,5-c]pyridine as a yellow solid (1.08g, yield: 81%). MS (ESI) calcd. for C₁₂H₇N₃O₂S (m/z): 257, found: 258[M+1]⁺.

Preparation of 3-(thiazolo[4,5-c]pyridin-2-yl)benzenamine

A mixture of 2-(3-nitrophenyl)thiazolo[4,5-c]pyridine (1.53 g, 5.9mmol), NH₄Cl (2.76 g, 51.6 mmol), iron powder (1.80 g, 32.2 mmol), H₂O(30 mL) and methanol (120 mL) was heated to reflux for 5.5 h under N₂.The mixture was filtered and the filtrate was diluted with H₂O (400 mL).The precipitate was collected by filtration and dried in vacuo to afford3-(thiazolo[4,5-c]pyridin-2-yl)benzenamine as a brown solid (841 mg,63%). ¹HNMR (400 MHz, DMSO-d₆) δ: 9.28 (1H, s); 8.52 (1H, s); 8.21 (1H,s); 7.23-7.38 (3H, d), 6.79 (1H, s), 5.51 (2H, s); MS (ESI) calcd. forC₁₂H₉N₃S (m/z): 227, found: 228 [M+H]P

General Procedure for Preparing Compounds 272, 273, 400, 401, 402, and403

Essentially the same procedure as detailed above for the preparation ofCompound 343 using 3-(thiazolo[4,5-c]pyridin-2-yl)benzenamine as thestarting material and the appropriate acid chloride.

Preparation of N-(4-hydroxypyridin-3-yl)-4-nitrobenzamide

A solution of p-nitrobenzoyl chloride (4.8 g, 25.7 mmol) in pyridine(77.0 mL) was added dropwise to a solution of 3-amino-4-hydroxypyridine(3.4 g, 30.9 mmol) in pyridine (51.0 mL) at 10° C. and stirredovernight. A solution of Na₂CO₃ (1.7 g) in water (65 mL) was added andthe resultant mixture was stirred for 1 h. The reaction mixture wasneutralized with 10% AcOH. The precipitate was collected by filtration,washed with 10% AcOH, and dried in vacuo (50° C.) to affordN-(4-hydroxypyridin-3-yl)-4-nitrobenzamide as a light green powder (4.1g, Yield: 62%). ¹HNMR (400 MHz, DMSO-d₆) δ: 11.64 (1H, br s), 9.57 (1H,s), 9.07 (1H, s), 8.75 (1H, d, J=4.4 Hz), 8.36 (1H, d, J=8.8 Hz), 8.20(1H, d, J=8.8 Hz), 7.74 (1H, d, J=4.4 Hz), 6.33 (1H, s); MS (ESI) calcdfor C₁₂H₉N₃O (m/z): 211, found: 212 [M+1]⁺.

Preparation of 2-(4-nitrophenyl)oxazolo[4,5-c]pyridine

A solution of N-(4-hydroxypyridin-3-yl)-4-nitrobenzamide (2.59 g, 10.0mmol) and polyphosphoric acid (20 mL) was stirred at 140° C. for 6 h.The reaction mixture was poured into distilled water (200 mL) and sodiumhydroxide was added until pH=5. The precipitate was collected byfiltration, washed with water until neutral and dried under vacuum toafford 2-(4-nitrophenyl)oxazolo[4,5-c]pyridine as a yellow solid (2.261g, Yield: 94%). MS (ESI) calcd. for C₁₂H₇NO₃ (m/z): 241, found: 242[M+1]⁺.

Preparation of 4-(oxazolo[4,5-c]pyridin-2-yl)benzenamine

2-(4-Nitrophenyl)oxazolo[4,5-c]pyridine (2.261 g, 9.3 mmol), iron powder(2.814 g, 48.0 mmol) and NH₄Cl (4.314 g, 80.1 mmol) were refluxed inCH₃OH/H₂O (4:1, 469 mL) for 6 h. The mixture was filtered and thefiltrate was evaporated in vacuo. The residue was purified bychromatography on silica gel eluted with petroleum ether:ethylacetate:Et₃N=160:40:1 to afford4-(oxazolo[4,5-c]pyridin-2-yl)benzenamine as a yellow solid (1.215 g,yield: 62%). ¹H NMR (400 MHz, DMSO-d₆) δ: 6.11 (2H, s), 6.69 (2H, d,J=8.4 Hz), 7.75 (1H, d, J=5.6 Hz), 7.87 (2H, d, J=8.4 Hz), 8.46 (1H, d,J=5.6 Hz), 8.94 (1H, s); MS (ESI) calcd. for C₁₂H₉N₃O (m/z): 211, found:212 [M+1]⁺.

General Procedure for Preparing Compounds 339, 340, 341, 342, 449, 410,411, and 412

Compounds 339, 340, 341, 342, 449, 410, 411, and 412 were prepared byessentially the same procedure as detailed above for the preparation ofCompound 343 using 4-(oxazolo[4,5-c]pyridin-2-yl)benzenamine as thestarting material and the appropriate acid chloride.

Preparation of 2-(4-nitrophenyl)thiazolo[4,5-c]pyridine

A mixture of N-(4-hydroxypyridin-3-yl)-4-nitrobenzamide (5.18 g, 0.02mol) and P₂S₅ (8.90 g, 0.04 mol) in pyridine (25 mL) and p-xylene (100mL) was stirred at 140° C. for 18 h. The solvent was removed under vacuowhile hot and the residue was purified by recrystallization from EtOH toafford 2-(4-nitrophenyl)thiazolo[4,5-c]pyridine as a yellow solid (3.00g, yield: 59%). MS (ESI), calcd for C₁₂H₇N₃O₂S (m/z): 257, found: 258[M+1]⁺.

Preparation of 4-(thiazolo[4,5-c]pyridin-2-yl)benzenamine

2-(4-nitrophenyl)thiazolo[4,5-c]pyridine (2.57 g, 0.01 mol), iron powder(2.8 g, 0.05 mmol) and NH₄Cl (4.32 g, 0.08 mol) were refluxed inCH₃OH/H₂O (4:1, 200 mL) for 6 h. The mixture was filtered and thefiltrate was evaporated in vacuo. The residue was washed with water (30ml) to give 4-(thiazolo[4,5-c]pyridin-2-yl)benzenamine as a white solid(1.37 g, Yield: 60%).

General Procedure for Preparing Compounds 322, 323, 324, 325, 409 and450

Compounds 322, 323, 324, 325, 409 and 450 were prepared by essentiallythe same procedure as detailed above for the preparation of Compound 343using 4-(oxazolo[4,5-c]pyridin-2-yl)benzenamine as the starting materialand the appropriate acid chloride.

Preparation of N-(4-hydroxypyridin-3-yl)-5-nitrothiophene-2-carboxamide

A mixture of 5-nitrothiophene-2-carboxylic acid (5.000 g, 28.9 mmol) inSOCl₂ (40 mL) was refluxed for 2 h. The excess SOCl₂ was evaporated invacuo and 3-amino-4-hydroxypyridine (2.65 g, 24.1 mmol) in pyridine (150mL) was added dropwise. The reaction mixture was stirred at 10° C.overnight. Na₂CO₃ (1.533 g, 14.5 mmol) in water (100 mL) was added andthe resultant mixture was stirred for additional 1 h. The precipitatewas collected by filtration, washed with 10% Na₂CO₃ (120 mL), and driedin vacuo to affordN-(4-hydroxypyridin-3-yl)-5-nitrothiophene-2-carboxamide as a yellowpowder (5.87 g, yield: 92%).

Preparation of 2-(5-nitrothiophen-2-yl)oxazolo[4,5-c]pyridine

A solution of N-(4-hydroxypyridin-3-yl)-5-nitrothiophene-2-carboxamide(2.385 g, 9.0 mmol) in polyphosphoric acid (18 mL) was stirred at 140°C. for 6 h. The reaction mixture was poured into distilled water andsodium hydroxide was added until pH=5. The precipitate was collected byfiltration, washed with water until neutral and dried to afford2-(5-nitrothiophen-2-yl)oxazolo[4,5-c]pyridine as a yellow solid (1.775g, yield: 80%). MS (ESI) calcd. for C₁₀H₅N₃O₃S (m/z): 247, found: 248[M+1]⁺.

Preparation of 5-(oxazolo[4,5-c]pyridin-2-yl)thiophen-2-amine

2-(5-nitrothiophen-2-yl)oxazolo[4,5-c]pyridine (1.775 g, 7.2 mmol), ironpowder (2.108 g, 36.0 mmol) and NH₄Cl (3.081 g, 57.6 mmol) were refluxedin CH₃OH/H₂O (4:1, 360 mL) for 6 h. The mixture was filtered and thefiltration was evaporated in vacuo. The residue was purified bychromatography on silica gel eluted with petroleum ether:ethylacetate:Et₃N=160:40:1 to afford5-(oxazolo[4,5-c]pyridin-2-yl)thiophen-2-amine as a yellow solid (1.1 g,yield: 70%). ¹HNMR (400 MHz, DMSO-d₆) δ: 8.83 (1H, s), 8.41 (1H, d,J=4.8 Hz), 7.67 (1H, d, J=4.8 Hz), 7.57 (1H, d, J=4.4 Hz), 6.91 (2H, s),6.03 (1H, d, J=4.4 Hz); MS (ESI) calcd. for C₁₀H₇N₃OS (m/z): 217, found:218 [M+1]⁺.

Preparation of Compounds 422, 423, 424, 425, 426, 427 and 428

Compounds 422, 423, 424, 425, 426, 427 and 428 were prepared byessentially the same procedure as detailed above for the preparation ofCompound 343 using 5-(oxazolo[4,5-c]pyridin-2-yl)thiophen-2-amine as thestarting material and the appropriate acid chloride.

Preparation of 2-(5-nitrothiophen-2-yl)thiazolo[4,5-c]pyridine

A mixture of compound 17 (1.97 g, 7.43 mmol) and P₂S₅ (3.30 g, 15 mmol)in pyridine (30 mL) and p-xylene (120 mL) was stirred at 140° C. for 18h. The solvent was removed in vacuo while hot and the residue waspurified by recrystallization from EtOH to afford compound 21 as ayellow solid (700 mg, yield: 35%, Lot#: MC0052-050-21). MS (ESI) calcd.for C₁₀H₅N₃O₂S₂ (m/z): 263, found: 264.1 [M+1]⁺.

Preparation of 5-(thiazolo[4,5-c]pyridin-2-yl)thiophen-2-amine

Compound 21 (700 mg, 2.66 mmol), iron powder (745 mg, 13.30 mmol) andNH₄Cl (1.36 g, 22 mmol) were refluxed in CH₃OH/H₂O (4:1, 150 mL) for 6h. The mixture was filtered and the filtrate was evaporated in vacuo.The residue was washed with water (30 mL) and dried to afford compound22 as a yellow solid (306 mg, yield: 50%).

Preparation of Compounds 515, 516, 517, 518, 519 and 520

Compounds 515, 516, 517, 518, 519 and 520 were prepared by essentiallythe same procedure as detailed above for the preparation of Compound 343using 5-(oxazolo[4,5-c]pyridin-2-yl)thiophen-2-amine as the startingmaterial and the appropriate acid chloride.

Preparation of N-(3-hydroxypyridin-4-yl)-3-methylbenzamide

3-Amino-4-hydroxypyridine was prepared according to a procedure detailedin Journal of Organic Chemistry (1995), p. 5721. A solution of3-nitrobenzoyl chloride (3.10 g, 16.7 mmol) in pyridine (50 mL) wasadded dropwise to a solution of 3-amino-4-hydroxypyridine (2.481 g, 21.6mmol) in pyridine (40 mL) at 10° C. and the resultant mixture wasstirred overnight. Na₂CO₃ (1 g) was added and the mixture was stirredfor additional 1 hr. The solid was collected by filtration, washed with10% acetic acid (30 mL×3) and water (30 mL×3) and dried under vacuo toafford N-(3-hydroxypyridin-4-yl)-3-methylbenzamide as yellow solid(1.057 g, yield: 43%).

Preparation of 2-(3-nitrophenyl)oxazolo[5,4-c]pyridine

A solution of N-(3-hydroxypyridin-4-yl)-3-methylbenzamide (1.30 g, 5mmol) in polyphosphoric acid (7.5 mL) was stirred at 150° C. for 6 hrs.The reaction mixture was poured into distilled water and sodiumhydroxide was added until pH=5. The precipitate was collected byfiltration, washed with water until neutral and dried in vacuo oven (50°C.) to afford 2-(3-nitrophenyl)oxazolo[5,4-c]pyridine as yellow solid(1.120 g, yield: 93%).

Preparation of 3-(oxazolo[5,4-c]pyridin-2-yl)benzenamine

A suspension of 2-(3-nitrophenyl)oxazolo[5,4-c]pyridine (1.20 g, 5mmol), iron powder (1.40 g, 25 mmol) and NH₄Cl (2.14 g, 40 mmol) inCH₃OH:H₂O (4:1, 60 mL) was refluxed for 6 hrs. The reaction mixture wasfiltered and the filtrate was evaporated in vacuo. The residue waspoured into water. The precipitate was collected by filtration, andwashed with water (20 mL×3) to afford3-(oxazolo[5,4-c]pyridin-2-yl)benzenamine as a white solid (0.330 g,yield: 31%).

Preparation of Compounds 429, 430, 431, 451, 452, 453 and 454

Compounds 429, 430, 431, 451, 452, 453 and 454 were prepared byessentially the same procedure as detailed above for the preparation ofCompound 343 using 3-(oxazolo[5,4-c]pyridin-2-yl)benzenamine as thestarting material and the appropriate acid chloride.

Preparation of N-(3-hydroxypyridin-4-yl)-4-nitrobenzamide

A solution of p-nitrobenzoyl chloride (4.8 g, 25.7 mmol) in pyridine (50mL) was added to a solution of 4-amino-3-hydroxypyridine (2.5 g, 22.7mmol) in pyridine (88 mL) at 10° C. and stirred at room temperatureovernight. To the reaction mixture was added Na₂CO₃ (1.1 g, 10.4 mmol)in water (10 mL) and stirred for 1 hrs. 10% AcOH was added to neutralizethe solution. The precipitate was collected by filtration, washed with10% AcOH and dried to give N-(3-hydroxypyridin-4-yl)-4-nitrobenzamide asa yellow powder (2.27 g, yield: 52%). MS (ESI) calcd. for C₁₂H₉N₃O₄(m/z): 259, found: 260 [M+H]⁺.

Preparation of 2-(4-nitrophenyl)oxazolo[5,4-c]pyridine

A mixture of N-(3-hydroxypyridin-4-yl)-4-nitrobenzamide (260 mg, 1 mmol)and polyphosphoric acid (1.5 mL) was stirred at 150° C. for 6 hrs. Thereaction mixture was poured into water and sodium hydroxide was addeduntil pH=5. The precipitate was collected by filtration, washed withwater until neutral and dried to give2-(4-nitrophenyl)oxazolo[5,4-c]pyridine as brown solid (177 mg, yield:73%). ¹HNMR (400 MHz, DMSO-d₆) δ: 9.21 (1H, s), 8.62 (1H, d, J=4.8 Hz),8.51 (4H, d, J=18, 8.8 Hz), 7.95 (1H, d, J=5.2 Hz).

Preparation of 4-(oxazolo[5,4-c]pyridin-2-yl)benzenamine

A mixture of 2-(4-nitrophenyl)oxazolo[5,4-c]pyridine (610 mg, 2.5 mmol),NH₄Cl (1.2 g, 22.4 mmol), iron powder (0.76 g, 13.6 mmol), H₂O (13 mL)and methanol (51 mL) was heated to reflux for 6 h under N₂. The mixturewas concentrated in vacuo and purified by chromatography on silica gel(eluted with EA/PE=3:1) to give4-(oxazolo[5,4-c]pyridin-2-yl)benzenamine as light brown powder (340 mg,yield 64%). ¹H NMR (400 MHz, DMSO-d₆) δ: 8.95 (1H, s); 8.46 (1H, d,J=5.2); 7.92 (2H, d, J=8.8); 7.69 (1H, d, J=5.6), 6.71 (2H, d, J=8.8),6.19 (2H, s); MS (ESI) calcd for C₁₂H₉N₃O (m/z): 211, found: 212 [M+H]⁺

Preparation of Compounds 408 and 419

Compounds 408 and 419 were prepared by essentially the same procedure asdetailed above for the preparation of Compound 343 using4-(oxazolo[5,4-c]pyridin-2-yl)benzenamine as the starting material andthe appropriate acid chloride.

Preparation of N-(3-hydroxypyridin-4-yl)-5-nitrothiophene-2-carboxamide

A solution of 5-nitrothiophene-2carboxylic acid (5.0 g, 28.9 mmol) andSOCl₂ (40 mL) was refluxed for 2 hrs. The solvent was removed in vacuofollowed by addition of 4-amino-3-hydroxypyridine (2.65 g, 24.1 mmol) inpyridine (150 mL). The mixture was stirred at 10° C. overnight. Na₂CO₃(1.53 g, 14.5 mmol) in water (100 mL) was added and stirred for 1 hrfollowed by adding AcOH to adjust pH=7. The precipitate was collected byfiltration, washed with 10% AcOH (30 mL×2) and dried to giveN-(3-hydroxypyridin-4-yl)-5-nitrothiophene-2-carboxamide as a yellowpowder (5.0 g, yield 78%).

Preparation of 2-(5-nitrothiophen-2-yl)oxazolo[5,4-c]pyridine

To a stirred solution ofN-(3-hydroxypyridin-4-yl)-5-nitrothiophene-2-carboxamide (1.1 g, 4.1mmol) in pyridine (5 mL) was added P₂O₅ (1.2 g, 8.3 mmol) and p-xylene(21 mL). After refluxing at 160° C. overnight, the reaction mixture wasconcentrated in vacuo and the residue was purified by chromatography onsilica gel eluted with 5% of ethyl acetate in CH₂Cl₂ to give2-(5-nitrothiophen-2-yl)oxazolo[5,4-c]pyridine as brown solid (174 mg,yield 17%). ¹H NMR (400 MHz, CDCl₃) δ: 8.97 (1H, s); 8.59 (1H, d,J=5.6); 7.94 (1H, d, J=4.4); 7.86 (1H, d, J=4.4), 7.69 (1H, d, J=5.6,0.8).

Preparation of 5-(oxazolo[5,4-c]pyridin-2-yl)thiophen-2-amine

A mixture of 2-(5-nitrothiophen-2-yl)oxazolo[5,4-c]pyridine (170 mg,0.69 mmol), NH₄Cl (199 mg, 3.73 mmol), iron powder (328 mg, 5.87 mmol),H₂O (10 mL) and methanol (40 mL) was heated to reflux for 6 hrs underN₂. The mixture was concentrated in vacuo followed by addition of 100 mLof water and left overnight at 4° C. The precipitate was collected byfiltration and dried to give5-(oxazolo[5,4-c]pyridin-2-yl)thiophen-2-amine as a brown solid (52 mg,yield 35%). ¹HNMR (400 MHz, DMSO-d₆) δ: 8.84 (1H, s); 8.40 (1H, d,J=5.2); 7.64 (1H, d, J=4); 7.57 (1H, d, J=5.2), 7.06 (2H, s), 1.09 (1H,d, J=4.4).

Preparation of Compounds 561, 562 and 563

Compounds 561, 562 and 563 were prepared by essentially the sameprocedure as detailed above for the preparation of Compound 343 using5-(oxazolo[5,4-c]pyridin-2-yl)thiophen-2-amine as the starting materialand the appropriate acid chloride.

Preparation of 2-(oxazolo[4,5-c]pyridin-2-yl)benzenamine

3-Amino-4-hydroxypyridine (2.225 g, 20.0 mmol) and 2-aminobenzoic acid(2.740 g, 20.0 mmol) were stirred in polyphosphoric acid (40 mL) at 140°C. for 6 hrs. The reaction mixture was poured into distilled water andsodium hydroxide was added until pH=8. The precipitate was collected byfiltration and the crude product was purified by chromatography onsilica gel eluting with petroleum ether:ethyl acetate:Et₃N (160:40:1) togive 2-(oxazolo[4,5-c]pyridin-2-yl)benzenamine (1.659 g, Yield: 39%). ¹HNMR (400 MHz, DMSO-d₆) δ: 6.68 (1H, t, J=7.6 Hz), 6.93 (1H, d, J=8.0Hz), 7.17 (2H, s), 7.30 (1H, t, J=7.6 Hz), 7.83 (1H, d, J=5.6 Hz), 7.90(1H, d, J=8.0 Hz), 8.55 (1H, d, J=5.6 Hz), 9.05 (1H, s); MS (ESI) calcd.for C₁₂H₉N₃O (m/z): 211, found: 212 [M+1]⁺.

Preparation of Compounds 404, 405, 406, 407, 420 and 421

Compounds 404, 405, 406, 407, 420 and 421 were prepared by essentiallythe same procedure as detailed above for the preparation of Compound 343using 2-(oxazolo[4,5-c]pyridin-2-yl)benzenamine as the starting materialand the appropriate acid chloride.

Preparation of 2-(2-nitrophenyl)thiazolo[4,5-c]pyridine

N-(4-Hydroxy-pyridin-3-yl)-2-nitro-benzamide was prepared using aprocedure similar to that described above using4-hydroxy-3-aminopyridine and 2-nitrobenzoyl chloride. A mixture of-(4-Hydroxy-pyridin-3-yl)-2-nitro-benzamide (2.6 g, 0.01 mol) and P₂S₅(4.44 g, 0.02 mol) in pyridine (12.5 mL) and p-xylene (50 mL) wasstirred at 140° C. for 18 hrs. The solvent was removed under vacuo andthe residue was purified by recrystallization to give2-(2-nitrophenyl)thiazolo[4,5-c]pyridine as a yellow solid (1.43 g,yield: 55%).

Preparation of 2-thiazolo[4,5-c]pyridin-2-yl-phenylamine

A suspension of 2-(2-nitrophenyl)thiazolo[4,5-c]pyridine (1.170 g, 4.6mmol), Fe powder (1.26 g, 22.8 mmol) and NH₄Cl (1.97 g, 36.8 mmol) inCH₃OH: H₂O (4:1, 80 mL) was refluxed for 6 hrs. The mixture was filteredand the filtrate was evaporated in vacuo. The residue was poured intowater. The precipitate was collected by filtration, and washed withwater (20 mL×3) to afford 2-thiazolo[4,5-c]pyridin-2-yl-phenylamine as ayellow solid (0.760 g, yield: 73%).

Preparation of Compounds 317, 318, 319, 320, 321 and 349

Compounds 317, 318, 319, 320, 321 and 349 were prepared by essentiallythe same procedure as detailed above for the preparation of Compound 343using 2-(2-nitrophenyl)thiazolo[4,5-c]pyridine as the starting materialand the appropriate acid chloride.

Preparation of 2-(oxazolo[5,4-c]pyridin-2-yl)benzenamine

3-Hydroxy-4-aminopyridine (2.220 g, 20.0 mmol) and 2-aminobenzoic acid(2.740 g, 20.0 mmol) in polyphosphoric acid (40 mL) were stirred at 140°C. for 6 hrs. The reaction mixture was poured into distilled water andsodium hydroxide was added until pH=8. The precipitate was collected byfiltration and further purified by chromatography on silica gel elutedwith petroleum ether:ethyl acetate:Et₃N (160:40:1) to give2-(oxazolo[5,4-c]pyridin-2-yl)benzenamine as yellow solid (1.332 g,Yield: 31%). ¹H NMR (400 MHz, DMSO-d₆) δ: 6.70 (1H, t, J=6.8 Hz), 6.94(1H, d, J=8.4 Hz), 7.20 (2H, s), 7.33 (1H, t, J=6.8 Hz), 7.80 (1H, d,J=5.6 Hz), 7.95 (1H, d, J=8.4 Hz), 8.53 (1H, d, J=5.6 Hz), 9.05 (1H, s);MS (ESI) calcd. for C₁₂H₉N₃O (m/z): 211, found: 212 [M+1]⁺.

Preparation of Compounds 455, 456, 457, 458 and 459

Compounds 455, 456, 457, 458 and 459 werer prepared by essentially thesame procedure as detailed above for the preparation of Compound 343using 2-(oxazolo[5,4-c]pyridin-2-yl)benzenamine as the starting materialand the appropriate acid chloride.

Preparation of N-(2-chloro-5-methylpyridin-3-yl)-2-nitrobenzamide

To a solution of 5-amino-6-chloro-3-picoline (9.54 g, 66.9 mmol) inpyridine (200 mL) was added 2-nitrobenzoyl chloride (13.65 g, 73.6 mmol)dropwise at 0° C. The resulting mixture was stirred at room temperaturefor 18 h. The dark mixture was then diluted with water (1500 mL) andsat. sodium bicarbonate solution was added until pH=8. The precipitatewas collected by filtration, rinsed with water (30 mL×3) and dried inoven to afford N-(2-chloro-5-methylpyridin-3-yl)-2-nitrobenzamide as apale solid (17.70 g, yield: 91%).

Preparation of 6-methyl-2-(2-nitrophenyl)thiazolo[5,4-b]pyridine

A mixture of N-(2-chloro-5-methylpyridin-3-yl)-2-nitrobenzamide (5.0 g,17.1 mmol) and P₂S₅ (7.6 g, 34.2 mmol) in pyridine (50 mL) and p-xylene(200 mL) was stirred at 140° C. for 20 h. The hot solution wastransferred to another flask and the solvent was removed in vacuo. Theresidue was purified by recrystallization from EtOH to afford6-methyl-2-(2-nitrophenyl)thiazolo[5,4-b]pyridine as a yellow solid (3.5g, yield: 75%).

Preparation of 6-(bromomethyl)-2-(2-nitrophenyl)thiazolo[5,4-b]pyridine

6-Methyl-2-(2-nitrophenyl)thiazolo[5,4-b]pyridine (2.9 g, 10.7 mmol),N-bromosuccinimide (NBS, 1.91 g, 10.7 mmol), CCl₄ (200 mL) and benzoylperoxide (0.021 g) were added into a three-neck flask (500 mL) underargon. The resulting yellow mixture was refluxed for 2 h. Additional NBS(1.91 g) and benzoyl peroxide (0.021 g) were added. Two hours later,more NBS (0.95 g) and benzoyl peroxide (0.021 g) were added and themixture was continually refluxed for 3 h. The mixture was then cooled toroom temperature. The solution was transferred into another flask andconcentrated in vacuo to afford crude6-(bromomethyl)-2-(2-nitrophenyl)thiazolo[5,4-b]pyridine (4.0 g), whichwas directly used for next step.

Preparation of tert-butyl4-((2-(2-nitrophenyl)thiazolo[5,4-b]pyridin-6-yl)methyl)piperazine-1-carboxylate

A solution of crude6-(bromomethyl)-2-(2-nitrophenyl)thiazolo[5,4-b]pyridine (4.0 g),Boc-piperazine (1.99 g, 10.7 mmol), Et₃N (1.5 mL, 10.7 mmol) andacetonitrile (100 mL) was stirred at 50° C. for 4 h and then at roomtemperature for 60 h. TLC demonstrated that the reaction was complete.The mixture was concentrated in vacuo and purified by silica gelchromatography (petroleum ether:ethyl acetate:Et₃N=100:10:1) to affordtert-butyl4-((2-(2-nitrophenyl)thiazolo[5,4-b]pyridin-6-yl)methyl)piperazine-1-carboxylateas a yellow solid (3.6 g, yield: 74% over 2 steps).

Preparation of tert-butyl4-((2-(2-aminophenyl)thiazolo[5,4-b]pyridin-6-yl)methyl)piperazine-1-carboxylate

A mixture of tert-butyl4-((2-(2-nitrophenyl)thiazolo[5,4-b]pyridin-6-yl)methyl)piperazine-1-carboxylate(2.13 g, 4.7 mmol), NH₄Cl (2.00 g, 37 mmol), iron powder (1.31 g, 23.5mmol), H₂O (40 mL) and methanol (160 mL) was refluxed for 3 h under N₂.The reaction mixture was filtered, and the filtrate was concentrated invacuo and purified by silica gel chromatography (petroleum ether:ethylacetate:Et₃N=800:200:1) to afford tert-butyl4-((2-(2-aminophenyl)thiazolo[5,4-b]pyridin-6-yl)methyl)piperazine-1-carboxylateas a yellow solid (1.63 g, yield: 81%).

General Procedure for Preparing Compounds 588, 589, 590, 591, 592, 593,594, 622, 646, 681, 682, 683, 684, 685, 686, 687, 688, 689, 701, 702,722, 723, 724, 725, 730, 731 and 732

A mixture of the amino scaffold (tert-butyl4-((2-(2-aminophenyl)thiazolo[5,4-b]pyridin-6-yl)methyl)piperazine-1-carboxylate,0.141 mmol each) and the appropriate acid chloride (0.17 mmol) inpyridine (2 mL) was shaken at room temperature overnight. The reactionmixture was then diluted with sat. NaHCO₃ (5 mL each). The precipitateswere collected by filtration and triturated with MeOH (5 mL) and driedto afford Boc-protected library compounds, which were analyzed by HPLC &MS. For those Boc-protected library compounds with purity below 95%, thecompounds were further purified by passing through silica gel pad elutedwith petroleum ether/EtOAc.

The Boc-protected compounds with were dissolved in 25% TFA/CH₂Cl₂solution (1 or 2 mL) and shaken at room temperature and monitored by TLCor LC/MS. The mixtures were then concentrated in vacuo and co-evaporatedwith CH₂Cl₂ (1.0 mL×2) in vacuo to afford the desired products as theTFA salts, which were analyzed by ¹H NMR, HPLC and MS.

Preparation of Compounds 690, 726, 727, 728 and 729

Compounds 690, 726, 727, 728 and 729 were prepared by essentially thesame procedure as detailed in the preparation of Compound 646 exceptthat 3-amino-2-hydroxy-6-methylpyridine was used as the startingmaterial.

General Procedure for Preparing Compounds 240, 608, 241, 221, 280, 222,223, 225, 244, 245, 246, 247, 226, 303, 238, 227, 304, 228, 305, 306,307, 308, 309, 310, 248, 249, and 396

Compounds 240, 608, 241, 221, 280, 222, 223, 225, 244, 245, 246, 247,226, 303, 238, 227, 304, 228, 305, 306, 307, 308, 309, 310, 248, 249,and 396 were prepared by essentially the same procedure as detailedbelow for the preparation of Compound 241 using3-Thiazolo[5,4-c]pyridin-2-yl-phenylamine as the starting material andthe appropriate acid chloride.

To a mixture of 3-thiazolo[5,4-c]pyridin-2-yl-phenylamine in 1.5 mL DMFwas added 5-phenyl furan-2-carbonyl chloride (62 mg, MW=206.63, 0.3mmol) in pyridine (0.25 mL). The reaction was kept for 72 hours at 40°C. and monitored with TLC. At the end of reaction, 5 mL H₂O was added,and the resulting suspension was filtered to collect the solid, whichwas then washed with H₂O and dried. The crude product was furtherpurified by preparative TLC (acetone/petroleum ether=1/2) and washedwith acetone to afford a white powder (43 mg, 36.1%). The product wasconfirmed by ¹H NMR. ¹H NMR (DMSO-d₆, 500 MHz), δ 10.50 (1H, s), 9.45(1H, s), 8.70 (1H, brs), 8.68 (1H, d, J=5.5 Hz), 8.13 (1H, d, J=7.8 Hz),8.08 (1H, d, J=5.5 Hz), 8.02 (2H, d, J=7.6 Hz), 7.93 (1H, d, J=7.7 Hz),7.63 (1H, t, J=7.9 Hz), 7.55 (3H, m), 7.43 (1H, t, J=7.3 Hz), 7.23 (1H,d, J=3.4 Hz). EIMS m/z (%): 397.2 (M⁺).

General Procedure for Preparing Compounds 466, 398, 229, 230, 282, 250,231, 399, 251, 283, 232, 252, 284, 285, 287, 234, 235, 236, 237, 239,289, 288, 290, 655, and 280

Compounds 466, 398, 229, 230, 282, 250, 231, 399, 251, 283, 232, 252,284, 285, 287, 234, 235, 236, 237, 239, 289, 288, 290, 655, and 280 wereby prepared by essentially the same procedure as detailed below for thepreparation of Compound 398 using3-Thiazolo[5,4-c]pyridin-2-yl-phenylamine as the starting material andthe appropriate sulfonyl chloride.

3-Thiazolo[5,4-c]pyridin-2-yl-phenylamine (68 mg, 0.3 mmol) in 2 mLpyridine was added to Dansyl chloride (81 mg, 0.3 mmol) while stirring.The reaction was kept at about 40° C. for 24 hours. Pyridine wasevaporated in vacuum, and water was added. The resulting suspension wasfiltered and washed by THF to afford the product as a yellow solid (29mg, 21.0%). ¹H NMR (DMSO-d₆, 500 MHz) δ 9.38 (1H, s), 8.64 (1H, d, J=2.8Hz), 8.45 (1H, d, J=8.5 Hz), 8.38 (1H, d, J=8.6 Hz), 8.30 (1H, d, J=7.1Hz), 8.03 (1H, d, J=5.4 Hz), 7.87(1H,s), 7.71(1H, d, J=7.6 Hz), 7.64(2H, m), 7.40 (1H, t, J=7.9 Hz), 7.31(1H, d, J=8.1 Hz), 7.26 (1H, d,J=7.6 Hz), 2.77 (6H, s). EIMS m/z: 460.01 (M⁺, 31).

General Procedure for Preparing Compounds 599, 600, 498, 610, 601, 611,485, 484, 481, 612, 475, 473, 472, 613, and 491

Compounds 599, 600, 498, 610, 601, 611, 485, 484, 481, 612, 475, 473,472, 613, and 491 were prepared by essentially the same procedure asdetailed below for the preparation of Compound 599 using3-Thiazolo[5,4-c]pyridin-2-yl-phenyl amine as the starting material andthe appropriate isothiocyanate.

To a mixture of 3-thiazolo[5,4-c]pyridin-2-yl-phenylamine in 1 mLpyridine was added 2,4-Dimethoxyphenyl Isothiocyanate (59 mg, 0.3 mmol).The reaction was kept for 24 hours at 60° C. with stirring and thereaction was monitored by TLC. Once completed, to the crude mixture wasadded 5 mL H₂O, and the resulting suspension was filtered to collect thesolid, which was then washed with H₂O and purified by preparative TLC(acetone/petroleum ether=1/2). The isolated solid was further washedwith acetone and dried in air to provide the desired product 38 mg(30.3%). The product was confirmed by ¹HNMR. ¹H NMR (DMSO-d₆, 500 MHz) δ9.40(1H, s), 8.65(1H, d, J=5.4 Hz), 8.55(1H, s), 8.05 (1H, d, J=5.2 Hz),7.90(1H, d, J=7.4 Hz), 7.77(1H, d, J=7.7 Hz), 7.55(1H, t, J=7.8 Hz),7.47(1H, d, J=6.6 Hz), 6.63 (1H, s), 6.54(1H, d, J=7.8 Hz), 3.82(3H, s),3.77(3H, s). MS: 422.26(M⁺).

General Procedure for Preparing Compounds 604, 605 and 607

Compounds 604, 605 and 607 were prepared by essentially the sameprocedure as detailed below for the preparation of Compound 604 using3-Thiazolo[5,4-c]pyridin-2-yl-phenylamine as the starting material andthe appropriate Chloroformate.

To a mixture of 3-thiazolo[5,4-c]pyridin-2-yl-phenylamine (68 mg, 0.3mmol) in 2.5 mL Pyridine was added to N-Methyl-N-phenylcarbamoylchloride (50.7 mg, 0.3 mmol). The reaction was kept at room temperaturefor 48 hours. To the mixture was then added water (5 mL) and theresulting suspension was filtered. The collected solid was furtherpurified by preparative TLC (petroleum ether/acetone=2/1). The productwas confirmed by ¹H NMR. ¹H NMR (500 MHz, Acetone-d₆) δ 9.34 (1H, s),8.66 (1H, d, J=5.3 Hz), 8.43 (1H, s), 7.97 (1H, d, J=5.4 Hz), 7.79 (2H,d, J=7.9 Hz), 7.74 (1H, d, J=8.1 Hz), 7.50 (2H, t, J=8.1 Hz), 7.44 (2H,d, J=8.0 Hz), 7.37 (1H, d, J=7.2 Hz), 3.36 (3H, s).

General Procedure for Preparing Compounds 387, 609, 390, 391, 462, 392,393, 436, 461, 460, 596, 465, and 463

Compounds 387, 609, 390, 391, 462, 392, 393, 436, 461, 460, 596, 465,and 463 were prepared by essentially the same procedure as detailedbelow for the preparation of Compound 385 using2-Thiazolo[5,4-c]pyridin-2-yl-phenylamine as the starting material andthe appropriate acid chloride.

To a mixture of 2-thiazolo[5,4-c]pyridin-2-yl-phenylamine in 1.5 mL DMFwas added 5-phenyl furan-2-carbonyl chloride (62 mg, 0.3 mmol) inpyridine (0.25 mL). The reaction was kept for 24 hours at 40° C. whilestirring and monitored by TLC. Once completed, to the crude mixture wasadded 5 mL H₂O, and the resulting suspension was filtered to collect thesolid product, which was subsequently washed with H₂O and dried. Thecrude product was further purified by preparative TLC (acetone/petroleumether=1/2) to yield 38 mg product (36.80%), confirmed by ¹H NMR: ¹H NMR(500 MHz, DMSO-d₆) δ 8.44 (1H, s), 7.74 (4H, d, J=7.5 Hz), 7.41 (4H, t,J=7.7 Hz), 7.29 (1H, t, J=7.4 Hz), 6.87 (2H, d, J=3.0 Hz), 6.75 (2H, d,J=2.6 Hz). EIMS m/z: 397.2 (M⁺).

Preparation of Compounds 162 and 163

A microwave vial was charged with2-thiazolo[5,4-c]pyridin-2-yl-phenylamine (40 mg, 0.2 mmol),3,4,5-trimethoxybenzoylchloride (40.6 mg, 0.2 mmol) and 1 mL ofpyridine. The mixture was subjected to microwave irradiation at 160° C.for 10 minutes. Upon cooling, MeOH was added to the solution, causing aprecipitate to form. The solid was filtered, washed with MeOH and driedto afford the desired amide product. (Calc'd for C21H17N3O3S: 391.45,[M+H]+ found: 392.1).

General Procedure for Preparing Compounds 467, 394 and 469

Compounds 467, 394 and 469 were prepared by essentially the sameprocedure as detailed below for the preparation of Compound 467 using2-Thiazolo[5,4-c]pyridin-2-yl-phenylamine as the starting material andthe appropriate sulfonyl chloride.

To a mixture of 2-thiazolo[5,4-c]pyridin-2-yl-phenylamine (71 mg, 0.3mmol) in 1 mL THF was added SM2 (68 mg, 0.3 mmol) in 2 mL pyridine withstirring at room temperature. The reaction was kept at 60° C. onoil-bath for 2 days and monitored via TLC till the starting materialdisappeared. 5 mL water was subsequently added and the resultingsuspension was filtered to collect the crude product, which was washedby acetone to yield a fine yellow powder 13 mg (10.2%).

¹H NMR (500 MHz, DMSO-d₆) δ 9.43 (1H, s), 8.70 (1H, d, J=5.5 Hz), 8.12(1H, d, J=5.4 Hz), 8.04 (1H, d, J=7.6 Hz), 7.58 (1H, t, J=7.5 Hz), 7.45(1H, d, J=8.1 Hz), 7.38 (1H, t, J=7.1 Hz), 7.17 (1H, d, J=8.3 Hz), 7.00(1H, s), 6.90 (1H, d, J=8.4 Hz), 3.71 (3H, s), 3.46 (3H, s). EIMS m/z:427.17 (M⁺, 10).

General Procedure for Preparing Compounds 614, 615 and 616

Compounds 614, 615 and 616 were prepared by essentially the sameprocedure as detailed below for the preparation of Compound 614 using2-Thiazolo[5,4-c]pyridin-2-yl-phenylamine as the starting material andthe appropriate isothiocyanate.

To a mixture of 2-Thiazolo[5,4-c]pyridin-2-yl-phenylamine (68 mg, 0.3mmol), 4-(1H-pyrazol-1-yl)phenyl isothiocyanate (60 mg, 0.3 mmol) and acatalytic amount of DMAP was added 8 mL Ethanol. The reaction was keptat 50° C. for 48 hours. The suspension was filtered, and washed withacetone thoroughly to yield 10 mg (7%) of product. ¹H NMR (500 MHz,DMSO-D6) δ 9.43 (1H, s), 8.58(1H, d, J=5.5 Hz), 8.47(1H, m), 8.21(1H, d,J=7.9 Hz), 8.19(1H, d, J=7.4 Hz), 7.85(2H, d, J=8.9 Hz), 7.80(1H, d,J=8.9 Hz), 7.75(1H, d, J=6.7 Hz), 7.68(2H, m), 7.61(1H, m), 7.46(1H, m),6.55(1H, m).

General Procedure for Preparing Compound 597

Compound 597 was prepared by essentially the same procedure as detailedbelow for the preparation of Compound 597 using2-Thiazolo[5,4-c]pyridin-2-yl-phenylamine as the starting material andthe appropriate Chloroformate.

4-Methoxyphenyl chloroformate (56 mg, 0.3 mmol) was added to2-Thiazolo[5,4-c]pyridin-2-yl-phenylamine (68 mg, 0.3 mmol) in 2.5 mlpyridine and the mixture was stirred at room temperature for 24 h. Water(5 mL) was added and the resulting mixture was filtered. The collectedcrude product was washed by water and dried to afford 11 mg (12%). ¹HNMR (500 MHz, Acetone-d6) δ 9.27 (1H, s), 8.73 (1H, d, J=5.5 Hz), 8.62(1H, d, J=8.4 Hz), 8.02(1H, d, J=5.5 Hz), 7.98(1H, d, J=7.8 Hz), 7.58(1H, m), 7.23 (3H, m), 6.97 (2H, d, J=8.9 Hz), 3.87 (3H, s).

Preparation of3-(5-dimethylaminomethyl-oxazolo[4,5-b]pyridin-2-yl)-phenylamine

5-Bromomethyl-2-(3-nitro-phenyl)-oxazolo[4,5-b]pyridine (250 mg, 0.75mmol) was dissolved in 5 mL of CH₃CN along 3 mL of a 2M solution ofdimethylamine in THF. The reaction mixture was stirred at roomtemperature for 18 hours. It was then concentrated and the resultingresidue was mixed with 25 mL of CH₂Cl₂ and washed with brine. Theorganic layer was dried (Na₂SO₄) and concentrated to afford essentiallyquantitative yield of the nitro intermediate. This was mixed with 6 mLof MeOH and 2 mL of water along with 200 mg of sodium hydrosulfidehydrate. The reaction mixture was stirred under reflux for 1 hour. Itwas then cooled to room temperature and concentrated to dryness. Theresulting residue was mixed with 100 mL of a 9:1 CH₂Cl₂/MeOH mixture andfiltered. The filtrate was concentrated to afford 120 mg of3-(5-dimethylaminomethyl-oxazolo[4,5-b]pyridin-2-yl)-phenylamine (MS,M⁺+H=269).

Preparation of Compound 165

3-(5-Dimethylaminomethyl-oxazolo[4,5-b]pyridin-2-yl)-phenylamine (54 mg,0.2 mmol) was reacted with 3-dimethylaminobenzoyl chloride in 1 mL ofpyridine using the same microwave conditions detailed earlier. Thereaction mixture was cooled to room temperature and concentrated.Purification by chromatography using a 9:1 mixture of CH₂Cl₂/MeOHbuffered with 1% triethylamine afforded 7 mg of the desired product (MS,M⁺+H=416).

Preparation of 3-(5-methyl-oxazolo[4,5-b]pyridin-2-yl)-phenylamine

5-Methyl-2-(3-nitro-phenyl)-oxazolo[4,5-b]pyridine (50 mg, 0.196 mmol)was mixed with 6 mL of MeOH along with 2 mL of water and 66 mg of sodiumhydrosulfide hydrate. The reaction mixture was stirred under reflux for2 hours. It was then concentrated and extracted with CH₂Cl₂. Thecombined organic layers were dried (Na₂SO₄) and concentrated to afford20 mg of 3-(5-methyl-oxazolo[4,5-b]pyridin-2-yl)-phenylamine.

Preparation of Compound 167

3-(5-Methyl-oxazolo[4,5-b]pyridin-2-yl)-phenylamine (20 mg, 0.0889 mmol)was reacted with 3-dimethylaminobenzoyl chloride in 1 mL of pyridineusing the same microwave conditions detailed earlier. The reactionmixture was cooled to room temperature and concentrated. Purification bychromatography using a 9:1 mixture of CH₂Cl₂/MeOH buffered with 1%triethylamine afforded 7 mg of the desired product (MS, M⁺+H=373).

Preparation of 5-(2-nitro-phenyl)-thiazol-2-ylamine

In a typical run, 2-amino-5-bromothiazole monohydrobromide (Aldrich,5.00 g, 0.0192 mol) was mixed with 40 mL of toluene, 40 mL of ethanol,and 20 mL of water. 2-Nitrophenyl boronic acid (3.2 g, 0.0192 mol) wasadded, along with 2.35 g of[1,1′-bis(diphenylphosphino)ferrocene]dichloro-palladium(II) complexwith CH₂Cl₂ (1:1) and 6.10 g of anhydrous sodium carbonate. The reactionmixture was stirred at 90° C. for 18 hours. It was then cooled to roomtemperature and concentrated. The resulting residue was mixed with 500mL of EtOAc and washed with water (3×50 mL). The organic layer wasfiltered to remove the black precipitate. The filtrate was extractedwith dilute 1N HCl. The combined aqueous layers were concentrated tonear dryness. The resulting residue was purified by preparative HPLCusing a mixture of aqueous acetonitrile that has been buffered with 0.1%TFA to afford 108 mg of 5-(2-nitro-phenyl)-thiazol-2-ylamine (MS,M⁺+H=222).

Preparation of 2-(2-nitro-phenyl)-imidazo[2,1-b]thiazole-6-carboxylicacid ethyl ester

5-(2-Nitro-phenyl)-thiazol-2-ylamine (100 mg, 0.452 mmol) was mixed with10 mL of methyl ethyl ketone along with 1.5 equivalents of ethylbromopyruvate. The reaction mixture was stirred under reflux for 5hours. The reaction mixture was cooled to room temperature andconcentrated. The crude product was purified by chromatography (Isco,gradient elution, CH₂Cl₂ to 9:1 CH₂Cl₂/MeOH) to afford 60 mg of2-(2-nitro-phenyl)-imidazo[2,1-b]thiazole-6-carboxylic acid ethyl ester(MS, M⁺+H=318).

Preparation of 2-(2-amino-phenyl)-imidazo[2,1-b]thiazole-6-carboxylicacid methyl ester

2-(2-Nitro-phenyl)-imidazo[2,1-b]thiazole-6-carboxylic acid ethyl ester(60 mg, 0.189 mmol) was mixed with 3 mL of MeOH along with sodiumhydrosulfide hydrate (32 mg, 0.567 mmol) in 1 mL of water. The reactionmixture was stirred under reflux for 1 hour and monitored by LC/MS.Reduction of the nitro group was complete at this point and the ethylester group had been exchanged to the corresponding methyl esterderivative. The reaction mixture was cooled to room temperature andconcentrated. The aqueous layer was extracted with CH₂Cl₂. The combinedorganic layers were dried (Na₂SO₄) and concentrated to affordessentially quantitative yield of2-(2-amino-phenyl)-imidazo[2,1-b]thiazole-6-carboxylic acid methyl ester(MS, M⁺+H=274).

Preparation of Compound 703

2-(2-Amino-phenyl)-imidazo[2,1-b]thiazole-6-carboxylic acid methyl ester(27 mg, 0.095 mmol) was mixed with 1 mL of pyridine along with 22 mg of3,4,5-trimethoxybenzoyl chloride. The reaction mixture was reacted in amicrowave reactor at 160° C. for 10 minutes. It was then cooled to roomtemperature and concentrated. The resulting crude product was purifiedby preparative HPLC using a mixture of aqueous acetonitrile that hasbeen buffered with 0.1% TFA to afford 108 mg of2-[2-(3,4,5-trimethoxybenzoylamino)-phenyl]-imidazo[2,1-b]thiazole-6-carboxylicacid methyl ester (MS, M⁺+H=468).

Preparation of Compound 704

2-[2-(3,4,5-trimethoxybenzoylamino)-phenyl]-imidazo[2,1-b]thiazole-6-carboxylicacid methyl ester (6 mg) was mixed with 1 mL of THF. Sodium hydroxide(10 mg) was added as a solution in 1 mL of water. The reaction mixturewas stirred at room temperature for 4 hours and then concentrated. Theresulting crude product was purified by preparative HPLC using a mixtureof aqueous acetonitrile that has been buffered with 0.1% TFA to afford108 mg of2-[2-(3,4,5-trimethoxy-benzoylamino)-phenyl]-imidazo[2,1b]thiazole-6-carboxylicacid (MS, M⁺+H=454).

Preparation of Compound 108

Tert-Butyl4-(3-amino-5-(oxazolo[4,5-b]pyridin-2-yl)benzoyl)piperazine-1-carboxylate(25 mg) was treated with 1 mL of a solution containing 25% TFA in CH₂Cl₂at room temperature for 30 min. It was then concentrated and Et₂O wasadded to precipitate out the product. After drying under vacuum, thedesired product was obtained as the TFA salt (MS, M⁺+H=324).

Preparation of 2-(methylthio)oxazolo[4,5-b]pyridine

2-(Methylthio)oxazolo[4,5-b]pyridine was prepared according to theprocedures of Chu-Moyer and Berger (J. Org. Chem. 1995, 60, 5721-5725)with minor modifications. To a suspension of 2-amino-3-hydroxypyridine(2.8 g, 25 mmol) in EtOH (62 mL, 0.4 M) was added potassium ethylxanthogenate (8.0 g, 50 mmol, 2 equiv). The reaction mixture was heatedto reflux (78° C.) and stirred for a period of 18 hours. The reactionmixture was concentrated to dryness and the resulting residue wasdissolved in water (70 mL). Upon acidification to pH 5 with glacialacetic acid, a large quantity of solid precipitated out. The suspensionwas filtered, washed with water (3×), and dried on the high-vac lineovernight to afford 2-thiooxazolo[4,5-b]pyridine as a brown solid (3.3g, 21.6 mmol, 86%).

To a cooled solution of 2-thiooxazolo[4,5-b]pyridine (3.3 g, 21.6 mmol)in DMF (54 mL, 0.4 M) at 0° C. was added K₂CO₃ (3.0 g, 21.6 mmol) andiodomethane (1.6 mL, 25.9 mmol). After stirring at 0° C. for 2.5 hours,the reaction mixture was diluted with water (60 mL) and extracted withEt₂O (3×100 mL). The combined organic layers were washed with water andbrine, dried (MgSO₄), filtered, and concentrated. Purification on silica(0% to 10% MeOH/CH₂Cl₂) afforded 2-(methylthio)oxazolo[4,5-b]pyridine asa tan-colored solid (2.5 g, 14.7 mmol, 68%). MS, [M+1]⁺=167.

Preparation of Boc-protected1-oxazolo[4,5-b]pyridin-2-yl-piperidin-3-ylamine

1-Oxazolo[4,5-b]pyridin-2-yl-piperidin-3-ylamine was prepared accordingto the procedures of Chu-Moyer and Berger (J. Org. Chem. 1995, 60,5721-5725) with minor modifications. To a solution of2-(methylthio)oxazolo[4,5-b]pyridine (1.9 g, 11.4 mmol) in toluene (1.1M) was added 3-Boc-piperidine (2.3 g, 11.4 mmol). The reaction mixturewas heated to 85° C. for 5.5 hours, cooled, then concentrated. Theresidue was dissolved in CH₂Cl₂, filtered, and concentrated.Purification on silica (0% to 10% MeOH/CH₂Cl₂) afforded the desiredproducts as a white solid (2.6 g, 8.3 mmol, 73%). MS, [M+1]⁺=319.

Preparation of Meta-Pyridyl 6-(1H-benzo[d]imidazol-2-yl)pyridin-2-amine

A suspension of 6-aminopyridine-2-carboxylic acid (1.12 g, 8.2 mmol)with 1,2-diaminobenzene (1.77 mg, 16.4 mmol) was heated in 8 mL of PPAat 180 degree for 2 h. The reaction mixture was poured into 250 mL ofice-water and neutralized with 2 N NaOH (chilled) while vigorouslystirred. The resulting precipitate was filtered to collect an off-whitesolid, which was washed with 20 mL warm water, dried and purified bychromatography using a 9:1 mixture of CH₂Cl₂ to MeOH (MS, M⁺+H=211).

Preparation of Compound 57

Meta-Pyridyl 6-(1H-benzo[d]imidazol-2-yl)pyridin-2-amine (25 mg, 0.118mmol) was mixed with 1 mL of pyridine along with 24 mg of2,3,4-trimethoxybenzoyl chloride (0.118 mmol). The reaction mixture wasreacted in a Biotage microwave reactor at 160° C. for 10 min. It wasthen cooled to room temperature and concentrated. The resulting crudeproduct was purified by chromatography (Isco, gradient elution, CH₂Cl₂to 9:1 CH₂Cl₂/MeOH) to afford 11 mg of the product (MS, M⁺+H=407.1).

Preparation of Compounds 58, 59, 60, 64, 69, 211 and 70

These compounds were prepared analogously to Compound 57, using theappropriate aromatic acid chlorides. The reaction mixture was eitherstirred at room temperature overnight or heated in a Biotage microwavereactor at 160 degrees for 10 minutes. It was then cooled to roomtemperature and concentrated under reduced pressure. The resultingresidual crude products were purified either by recrystallization usingacetonitrile or chromatography (Isco, gradient elution, CH₂Cl₂ to 9:1CH₂Cl₂/MeOH).

Preparation of Compound 71

In a 2 mL vial, 6-(oxazolo[4,5-b]pyridin-2-yl)pyridin-2-amine (11 mg,0.05 mmol) in 1 mL pyridine was added to5-isocyanatobenzo[d][1,3]dioxole (9 mg, 0.05 mmol). The reaction washeated to 160° C. (MW) for 10 min. An aliquot was taken and diluted withMeOH. The pyridine was removed under vacuum. The crude product wassuspended and magnetically stirred in 5 mL acetonitrile for half an hourand filtered to collect the product, which was subsequently dried undervacuum (MS, M⁺+H=376.1).

Preparation of Compounds 72, 87 and 147

These compounds were prepared analogously to Compound 71, using theappropriate aromatic isocyanates or isothiocyanates. The reactionmixture was either stirred at room temperature overnight or heated in aBiotage microwave reactor at 160 degree for 10 minutes. It was thencooled to room temperature and concentrated under reduced pressure. Theresulting residual crude products were purified either byrecrystallization using acetonitrile or chromatography (Isco, gradientelution, CH₂Cl₂ to 9:1 CH₂Cl₂/MeOH).

Preparation of 2-(1,4-diazepan-1-yl)benzo[d]oxazole

In a 40 mL vial at room temperature, 2-chlorobenzo[d]oxazole (760 mg, 5mmol) was added to a solution of homopiperazine (2 g, 20 mmol) in CH₃CN.The reaction mixture gradually turned into a suspension. Upon standingfor additional half an hour, solid was filtered off and the collectedfiltrate was directly subjected to chromatography (Isco, gradientelution, CH₂Cl₂ to 9:1 CH₂Cl₂/MeOH). 420 mg of the desired product wasisolated (MS, M⁺+H=218.1).

Preparation of Compound 85

Compound 85 was obtained from the above reaction to prepare2-(1,4-diazepan-1-yl)benzo[d]oxazole as a by product. 24 mg of productwas obtained (MS, M⁺+H=335.1).

Preparation of Compound 88

In a 2 mL vial, 2-(1,4-diazepan-1-yl)benzo[d]oxazole (43 mg, 0.2 mmol)in 1 mL pyridine was added 2,4-dimethoxybenzoyl chloride (40 mg,mw=230.6, 0.2 mmol). The reaction was heated to 160° C. (MW) for 10 minIt was then cooled to room temperature and concentrated. The resultingcrude product was purified by chromatography (Isco, gradient elution,CH₂Cl₂ to 9:1 CH₂Cl₂/MeOH) to afford 60 mg of the product (MS,M⁺+H=382.1).

Preparation of Compounds 89, 90 and 91

These compounds were prepared analogously to Compound 88, using theappropriate aromatic acid chlorides and isocyanate. The reaction mixturewas either stirred at room temperature overnight or heated in a Biotagemicrowave reactor at 160 degree for 10 minutes. It was then cooled toroom temperature and concentrated under reduced pressure. The resultingresidual crude products were purified either by recrystallization usingacetonitrile or chromatography (Isco, gradient elution, CH₂Cl₂ to 9:1CH₂Cl₂/MeOH)

Preparation of Compound 92

In a 2 mL vial, 3-(2-methylthiazol-4-yl)benzenamine (95 mg, 0.5 mmol,1.5 eq) was added to 2-chlorobenzo[d]oxazole (51 mg, 0.33 mmol) in 2 mLCH₃CN. The reaction was heated to 160° C. (MW) for 10 min. It was thencooled to room temperature and concentrated. The resulting crude productwas purified by chromatography (Isco, gradient elution, CH₂Cl₂ to 9:1CH₂Cl₂/MeOH) to afford 71 mg of the product (MS, M⁺+H=308.1).

Preparation of Compounds 93, 94, 95, 104 and 105

These compounds were prepared analogously to Compound 92 using3-(2-methylthiazol-4-yl)benzenamine, or6-(oxazolo[4,5-b]pyridin-2-yl)pyridin-2-amine, or6-(oxazolo[4,5-b]pyridin-2-yl)pyridin-2-amine to react with2-chlorobenzo[d]oxazole or 2-chlorobenzo[d]thiazole. The reactionmixture was heated in a Biotage microwave reactor at 160 degree for 10minutes. It was then cooled to room temperature and concentrated underreduced pressure. The resulting residual crude products were purifiedeither by recrystallization using acetonitrile or chromatography (Isco,gradient elution, CH₂Cl₂ to 9:1 CH₂Cl₂/MeOH).

Preparation of Compound 142

In a typical run, 2-(1H-benzo[d]imidazol-2-yl)benzenamine (105 mg, 0.5mmol) in 5 mL pyridine was added to 3,4-dimethoxybenzoyl chloride (100mg, mw=200, 0.5 mmol). The reaction was kept at room temperatureovernight while stirring. Pyridine was then removed and 5 mL MeOH wasthen added to the crude mixture. The resulting suspension, upon stirringfor half an hour, was filtered to collect off white solid, which wasthen washed with MeOH and dried under reduced pressure. TLC/HPLC/LC-Masssuggested that the purity of the product is greater than 95% (MS,M⁺+H=374.1).

Preparation of 3-(1H-benzo[d]imidazol-2-yl)pyridin-2-amine

A suspension of 2-aminopyridine-3-carboxylic acid 2-amino nicotinic acid(1.12 g, 8.2 mmol) with 1,2-diaminobenzene (1.77 mg, 16.4 mmol) washeated in 8 mL of PPA at 180 degree for 2 h. The reaction mixture waspoured into 250 mL ice-water and neutralized with 2 N ice-cold NaOHduring vigorous stirring. The resulting precipitate was filtered andwashed with 20 mL warm water to afford 1.1 grams of pure corresponding3-(1H-benzo[d]imidazol-2-yl)pyridin-2-amine (MS, M⁺+H=211.1).

Preparation of 6-(1H-benzo[d]imidazol-2-yl)pyridin-2-amine

A suspension of 6-aminopyridine-2-carboxylic acid (112 mg), or2-aminopyridine-4-carboxylic acid (112 mg, 0.82 mmol), or5-aminopyridine-3-carboxylic acid (112 mg, 0.82 mmol) with1,2-diaminobenzene (177 mg, 1.64 mmol) was heated in 4 mL of PPA (180degree) for 2 h. The reaction mixture was each poured into 50 mLice-water and neutralized with 2 N NaOH (chilled) during vigorousstirring. The resulting precipitate was filtered and washed with 20 mLwarm water to afford 105 mg of desired6-(1H-benzo[d]imidazol-2-yl)pyridin-2-amine (MS, M⁺+H=211.1).

Preparation of 3-(1H-benzo[d]imidazol-2-yl)pyridin-4-amine

A suspension of 4-aminopyridine-3-carboxylic acid 4-amino nicotinic acid(1.12 g, 8.2 mmol) with 1,2-diaminobenzene (1.77 mg, 16.4 mmol) washeated in 8 mL of PPA at 180 degrees for 2 h. The reaction mixture waspoured into 250 mL ice-water and neutralized with 2 N ice-cold NaOHduring vigorous stirring. The resulting precipitate was filtered andwashed with 20 mL warm water to afford 1.35 grams of pure corresponding3-(1H-benzo[d]imidazol-2-yl)pyridin-2-amine (MS, M⁺+H=211.1).

Preparation of 2-(1-methyl-1H-benzo[d]imidazol-2-yl)benzenamine

2-(1H-benzo[d]imidazol-2-yl)benzenamine (525 mg, 2.5 mmol) in 5 mL dryTHF was cooled to −78 degrees before 1 mL n-Bu-Li (2.5 mmol, 2.5 N inhexanes) was added via a syringe. The reaction was stirred at −78degrees for 20 min and MeI (360 mg, 2.6 mmol) was added. The reactionwas warmed to r.t. gradually and kept at room temperature for anotherhour. The reaction crude was diluted with 25 mL ether and the resultedsuspension was filtered off the solid. The crude product was isolatedafter the solvent was removed from the collected filtrate to product 510mg of 2-(1-methyl-1H-benzo[d]imidazol-2-yl)benzenamine in greater than95% purity (MS, M⁺+H=224.1).

Preparation of Compounds 141, 143, 144, 145, 146, 168, 169, 175, 176,177, 257, 258, 259, 260, 261, 276, 313, 314, 315, 507, 508, 556, and 293

These compounds were prepared analogously to Compound 142, using theappropriate substituted bicyclic aromatic benzenamines to react withcorresponding aromatic acid chlorides, sulfonyl chlorides,chloroformates and isocyanate. The reaction mixture was either stirredat room temperature overnight or heated in a Biotage microwave reactorat 160 degree for 10 minutes. It was then cooled to room temperature andconcentrated under reduced pressure. The resulting residual crudeproducts were purified either by recrystallization using acetonitrile orchromatography using a 9:1 mixture of CH₂Cl₂ to MeOH.

Preparation of Compound 503

To 3-methoxy-4-(morpholinomethyl)benzoic acid (502 mg, 2 mmol) in CH₂Cl₂(20 mL) was added (COCl)₂ (2 mL, 23 mmol), followed by one tiny drop ofDMF. The resulting mixture was stirred at room temperature for 3 h, andevaporated in vacuo to give a yellow solid. This solid was thendissolved in 5 mL DMF. 2-(1H-benzo[d]imidazol-2-yl)benzenamine (420 mg,2 mmol) in 5 mL pyridine was subsequently added and the reaction mixturewas kept at room temperature overnight while stirring. The reaction wasconcentrated to provide the crude product, which was further purified byShimadzu reverse prep HPLC to afford the desired product (103 mg)with >98% purity (MS, M⁺+H=443.1).

Preparation of Compound 587

3-methoxy-4-((pyrrolidin-1-yl)methyl)benzoate (249 mg, 1 mmol) and2-(1H-benzo[d]imidazol-2-yl)benzenamine (209 mg, 1 mmol) in toluene (5mL) was cooled to 0° C. and then AlMe₃ was added (2 mL, 2 M in toluene,4 eq). The resulting mixture was gradually warmed up to rt and stirredovernight. The reaction was then cooled again to 0° C. and carefullyquenched with 5 mL MeOH. After warmed up to room temperature, themixture was partitioned between saturated NaHCO₃ and AcOEt. Before theseparation of the two layers, the mixture was filtered. Then, thebiphasic filtrate was separated. The organic layer was collected anddried over Na₂SO₄, filtered and concentrated under reduced pressure. Theproduct was purified by combiflash using amine RediSep column(68-2203-100, Teledyne Isco) (0-2% MeOH in DCM). The collected fractionswere combined and concentrated to provide 25 mg of product (MS,M⁺+H=427.1).

Preparation of Compounds 557 and 558

These compounds were prepared analogously to Compound 587. The productwas purified by combiflash using amine RediSep columns.

Preparation of 3-(chloromethyl)-6-(2-nitrophenyl)imidazo[2,1-b]thiazole

(3-Nitro-5-thiazolo[5,4-c]pyridin-2-yl-phenyl)-methanol (1.375 g, 5mmol) was suspended in 25 mL of CH₂Cl₂ and cooled with an ice bath.Thionyl Chloride (3.6 mL, 10 eq) was added dropwise and the reactionmixture was slowly warmed to room temperature. After stirring overnight,to the reaction mixture was added 100 mL ether and the resultingsuspension was filtered to collect 1.55 g of the desired product (MS,M⁺+H=293.1).

Preparation of Compound 626

3-(chloromethyl)-6-(2-nitrophenyl)imidazo[2,1-b]thiazole (292 mg, 1mmol), 4-benzylpiperazin-2-one (380 mg, 2 mmol) and NaH (88 mg, 2.2 eq)were suspended in 4 mL of dry DMF. The reaction was heated overnight at100° C. After cooling to room temperature, the reaction mixture waspartitioned between water and AcOEt. The organic layer was collected,dried and evaporated to provide crude product, which was furtherpurified by reverse HPLC to give 211 mg of desired product4-benzyl-1-((6-(2-nitrophenyl)imidazo[2,1-b]thiazol-3-yl)methyl)piperazin-2-one(MS, M⁺+H=448.1).

Preparation of Compound 627

To a suspension of 200 mg of1-((6-(2-nitrophenyl)imidazo[2,1-b]thiazol-3-yl)methyl)-4-benzylpiperazin-2-onein 5 mL MeOH was added 250 mg of sodium hydrosulfide hydrate. Thereaction mixture was heated for 30 minutes at 135° C. (MW). Aftercooling to room temperature, the mixture was diluted with 50 mL of waterand extracted with CH₂Cl₂. The combined organic layers were dried(Na₂SO₄) and concentrated to afford the crude product, which can befurther purified via reverse phase HPLC to provide 135 mg of thetargeted product1-((6-(2-aminophenyl)imidazo[2,1-b]thiazol-3-yl)methyl)-4-benzylpiperazin-2-one(MS, M⁺+H=418.1).

Preparation of Compound 628

A mixture of1-((6-(2-aminophenyl)imidazo[2,1-b]thiazol-3-yl)methyl)-4-benzylpiperazin-2-one(44 mg, 0.1 mmol) and 2-quinoxaloyl chloride (21 mg, 1.1 eq) in 2.5 mLpyridine was heated for 20 minutes at 160° C. (MW). After cooling toroom temperature, pyridine was removed and the reaction crude wasredissolved in methanol and purified via reverse phase HPLC to provide22 mg desired product (MS, M⁺+H=574.1).

Preparation of 617, 618, 647, 648, 676, 677, 678, 679, 699, 741, 742 and711

These compounds were prepared analogously to Compound 628. Products werepurified via reverse phase HPLC.

Preparation of Compound 42

A 50 mL flask was charged with 2-amino-3-hydroxypyridine (1.0 g, 9.1mmol), 3-aminobenzoic acid (1.24 g, 9.1 mmol) and 12 mL ofpolyphosphoric acid. The mixture was brought to 200° C. After 4 h, thereaction was cooled slightly and then quenched with ice-water. Themixture was neutralized to pH 7.5 with sat. Na₂CO₃.

The product was extracted with EtOAc, and the combined organic extractswere washed with brine and dried over MgSO₄. Solvent evaporationafforded the desired aminophenyl-oxaolopyridine product. (Calc'd forC12H9N3O: 211.2, [M+H]+ found: 212)

To a solution of the aniline (0.375 g, 1.8 mmol) in 6 mL of DMF wasadded 3-dimethylaminobenzoic acid (0.29 g, 1.8 mmol), HATU (1.0 g, 2.7mmol), HOAt (0.36 g, 2.7 mmol) and diisopropylethylamine (0.69 g, 5.3mmol). The reaction mixture was stirred overnight at 70° C. Afterdilution with CH₂Cl₂, the organic layer was washed with sat. NaHCO₃ andbrine, and dried over MgSO₄. The crude material was purified by silicachromatography (0-10% MeOH/CH₂Cl₂) to afford the desired product.(Calc'd for C21H18N4O2: 358, [M+H]+ found: 359)

Compounds 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33,34, 35, 36, 37, 38, 39, 40, 41, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52,53, 54, 55, 56, 109 and 110 were prepared in an analogous manner toCompound 42.

Preparation of Compound 65

A microwave vial was charged with 4-oxazolo[4,5-b]pyridin-2-ylamine (50mg, 0.2 mmol), 4-methoxybenzensulfonyl chloride (49 mg, 0.2 mmol) and 1mL of pyridine.

The mixture was subjected to microwave irradiation at 160° C. for 12minutes. The pyridine was then evaporated, and the resulting brown solidwas triturated with methanol/dichloromethane. The solid was filtered,washed with methanol/dichloromethane and dried to afford the desiredproduct as an off-white powder. (Calc'd for C18H14N4O4S: 382.40, [M+H]+found: 383.0).

Compounds 61, 63, 75, 76, 96, 97, 98, and 106 were prepared in ananalogous manner to Compound 65.

Preparation Compound 77

A 50 mL round bottom flask was charged with 2-aminophenol (0.24 g, 2.2mmol), 2-aminopyridine-4-carboxylic acid (0.30 g, 2.2 mmol) and 2 mL ofpolyphosphoric acid. The mixture was brought to 200° C. and was stirredfor 4 hours. The reaction was then quenched with ice and basified to pH7.5 with saturated aqueous Na₂CO₃. The resulting precipitate wasfiltered, washed with water and dried to afford the desired product as abrown solid. (Calc'd for C12H9N3O: 211.23, [M+H]+ found: 212.1).

A microwave vial was charged with 4-benzoxazol-2-yl-pyridin-2-ylamine(0.050 g, 0.2 mmol), 3,4-dimethoxybenzoyl chloride (0.047 g, 0.2 mmol)and 1 mL of pyridine. The mixture was subjected to microwave irradiationat 160 oC for 12 minutes. Upon cooling, methanol was added to themixture and a precipitate formed. The solid was filtered, washed withmethanol and dried to afford the desired product as a red solid. (Calc'dfor C21H17N3O4: 375.39, [M+H]+ found: 376.1)

Compound 78 was prepared in an analogous manner to Compound 77.

Preparation of1-Benzo[1,3]dioxol-5-yl-3-(4-benzoxazol-2-yl-pyridin-2-yl)-urea;Compound 86

A microwave vial was charged with 4-benzoxazol-2-yl-pyridin-2-ylamine(0.05 g, 0.2 mmol), 3,4-(methylenedioxy)phenyl isocyanate (0.04 g, 0.2mmol) and 1 mL of pyridine. The mixture was subjected to microwaveirradiation at 160° C. for 15 minutes. Upon cooling, a precipitateformed. The solid was filtered and then triturated with hot methanol.The undissolved material was filtered, and the precipitate that formedin the mother liquor was filtered and dried to afford the desiredproduct as a white solid. (Calc'd for C20H14N4O4: 374.36, [M+H]+ found:375.1).

Preparation of2-[1-(4-Fluoro-phenyl)-5-methyl-1H-imidazol-4-yl]-oxazolo[4,5-b]pyridine;Compound 62

A 25 mL round bottom flask was charged with1-(4-fluorophenyl)-5-(trifluoromethyl)pyrazole carboxylic acid (0.15 g,0.5 mmol), 2-amino-3-hydroxypyridine (0.060 g, 0.5 mmol) and 1 mL ofpolyphosphoric acid. The mixture was brought to 200° C. and was stirredfor 4 hours. The reaction was then quenched with ice and basified to pH7.5 with saturated aqueous Na₂CO₃.

The resulting precipitate was filtered and washed with methanol. Thecrude product was purified by silica preparatory tlc plates, eluent 5%methanol/dichloromethane. The desired product was isolated and dried toafford a white powder. (Calc'd for C16H8F4N4O: 348.26, [M+H]+ found:349.0).

Preparation of3,4,5-Trimethoxy-N-methyl-N-(2-oxazolo[4,5-b]pyridin-2-yl-phenyl)-benzamide;Compound 292

To a suspension of3,4,5-trimethoxy-N-(2-oxazolo[4,5-b]pyridin-2-yl-phenyl)-benzamide (0.03g, 0.1 mmol) in 3 mL of DMF was added sodium hydride (60% dispersion inmineral oil, 0.006 g, 0.2 mmol). The mixture was stirred at RT for 30minutes, becoming homogeneous. To this was then added iodomethane, dropwise (0.040 g, 0.3 mmol, 0.02 mL). After an additional hour at RT, thereaction was quenched with water and then partitioned between saturatedNH₄Cl and EtOAc. The organic layer was washed with brine and dried overMgSO₄. The crude product was purified by silica chromatography, 0-5%methanol/dichloromethane. The desired product was isolated as a yellowsolid. (Calc'd for C23H21N3O5: 419.44, [M+H]+ found: 420.1)

Preparation of Compound 148

Para-nitrobenzoic acid (4.6 g, 27.5 mmol) and 2,3 diaminopyridine (3.0g, 27.5 mmol) were added to polyphosphoric acid (27.5 mL) at roomtemperature with a mechanical stirrer. The reaction mixture was stirredat 175° C. for 2 h, cooled to rt, quenched with water and basified withNaHCO₃. The aqueous layer was left to precipitate overnight at rt. Theresulting precipitate was filtered, washed with water, EtOAc and Et₂Oand dried in vacuo to give the desired product as a brown solid. (Calc'dfor C12H8N4O2: 240.22, [M+H]+ found: 241).

Dimethylformamide (30 mL) was added at 0° C. to sodium hydride (550 mg,13.7 mmol) prewashed with hexane under an Argon atmosphere.2-(4-Nitrophenyl)-3H-imidazo[4,5-b]pyridine (3.0 g, 12.43 mmol) wasadded portion-wise to this suspension at 0° C., and the reaction mixturewas stirred at rt for 1 h 40 min. Then2-(chloromethoxy)ethyltrimethylsilane (2.4 mL, 13.7 mmol) was added, andthe reaction mixture was stirred at rt for 2.5 h and quenched withwater. The aqueous layer was extracted with EtOAc and the combinedorganic layer was dried over Na₂SO₄, filtered and concentrated in vacuo.The crude product was purified by column chromatography on SiO2 withhexane/EtOAc (50:50) to (20:80) to give one fraction of pure isomer, anda second fraction which consisted of an 85:15 mixture of the tworegioisomers. (Calc'd for C18H22N4O3Si: 370.48, [M+H]+ found: 371).

Palladium(0) on charcoal (10% w/w, 160 mg, 0.15 mmol) was added at rt toa mixture of the SEM-protected imidazopyridine (1.60 g, 4.30 mmol) inMeOH (8 mL), EtOAc (12 mL) and methoxyethanol (2 mL). The reactionmixture was placed under an H₂-atmosphere (vacuum/Argon three times;vacuum/H₂ three times) and stirred at rt until complete conversion (4h). The reaction mixture was filtered over Celite and the filtrate wasconcentrated in vacuo to give the aniline as a brown solid. (Calc'd forC18H24N4OSi: 340.49, [M+H]+ found: 341).

To a solution of the imidazopyridine aniline (150 mg, 0.44 mmol) in 1.5mL of DMF was added 2,3,4-trimethoxybenzoic acid (93 mg, 0.44 mmol),HATU (250 mg, 0.7 mmol), HOAt (90 mg, 0.7 mmol) anddiisopropylethylamine (171 mg, 0.23 mL, 1.3 mmol). The reaction mixturewas stirred overnight at 80° C. After dilution with ETOAc, the organiclayer was washed with water, dried over Na₂SO₄, filtered andconcentrated in vacuo. The obtained compound was then dissolved in EtOH(1 mL) and 5 N aq. HCl (1 mL). The reaction mixture was stirred at 70°C. for 2.5 h, cooled to rt and neutralized with 2N aq. NaOH and sat. aq.NaHCO₃ solution. The mixture was partially concentrated in vacuo and theresulting precipitate was filtered, washed with water and Et₂O and driedunder vacuum to give the desired product.

Preparation of Compound 149, 150, 151, 180, 181, 182, 183, 184 and 185were done in an analogous manner to Compound 148.

Preparation of Compound 190

4-Ethynylaniline 15 (1.61 g, 13.79 mmol) was added at rt to a solutionof 3-bromo-2-pyridinone 33 (2.0 g, 11.49 mmol), PdCl₂(PPh₃)₂ (403 mg,0.574 mmol) and CuI (109 mg, 0.572 mmol) in degassed triethylamine (60ml). The reaction mixture was degassed for 5 min, stirred at 100° C. for6 h and concentrated in vacuo. The black residue was mixed withCH₂Cl₂/MeOH, filtered and concentrated in vacuo. The crude product waspurified by column chromatography on SiO2 with CH₂Cl₂/MeOH (100:0) to(99:1) to afford the desired product. (Calc'd for C13H10N2O: 210.23,[M+H]+ found: 211).

To a solution of theN-(4-furo[2,3-b]pyridine-2-yl-phenyl-2,3-dimethoxybenzamide (30 mg,0.143 mmol) and triethylamine (0.02 mL, 0.143 mmol) in CH₂Cl₂ (1 mL) wasadded 2,4-dimethoxybenzoyl chloride (44 mg, 0.219 mmol). The reactionmixture was stirred at rt overnight. After dilution with water, the aq.layer was extracted with CH₂Cl₂, and the combined organic layer wasdried over Na₂SO₄, filtered and concentrated in vacuo. The crude productwas purified by column chromatography on SiO₂ with Hexane/EtOAc (80:20to 50:50) to afford the desired product. (Calc'd for C22H18N2O4: 374.39,[M+H]+ found: 375).

Preparation of Compounds 186, 187, 188, 189, 191, 192 and 193 was donein an analogous manner as Compound 190.

Preparation of Compound 200

2-(3-Nitrophenyl)-thiazolo[4,5-b]pyridine was prepared from2-amino-pyridine in six steps using the procedure described in SulfurLetters 1995, 18 (2), 79-95. Palladium(0) on charcoal (10% w/w, 40 mg,0.038 mmol) was added at rt to a mixture of2-(3-nitrophenyl)-thiazolo[4,5-b]pyridine (403 mg, 1.57 mmol), toluene(2.5 ml), AcOH (2.5 ml) and methoxyethanol (2.5 ml). The reactionmixture was placed under an H2-atmosphere (vacuum/Argon three times;vacuum/H2 three times) and stirred at 50° C. for 3 h and at 60° C. for1.5 h. As the reaction was not completed, another portion of palladiumon charcoal (40 mg, 0.038 mmol) was added, and the reaction mixture wasstirred at 60° C. for 20 h. The reaction mixture was cooled to rt,filtered over Celite, and the filtrate was concentrated in vacuo. Thecrude product was purified by column chromatography on SiO2 withEtOAc/hexane (80:20) to give the desired aniline as a brown solid.(Calc'd for C12H9N3S: 227.27, [M+H]+ found: 228.2).

To a solution of the thiazolo[4.5-b]pyridin-2-yl-phenylamine (40 mg,0.176 mmol) and triethylamine (0.02 mL, 0.143 mmol) in CH₂Cl₂ (1 mL) wasadded 3-dimethylaminobenzoyl chloride hydrochloride (47 mg, 0.213 mmol).The reaction mixture was stirred at rt overnight. After dilution withwater, the aq. layer was extracted with CH₂Cl₂, and the combined organiclayer was dried over Na₂SO₄, filtered and concentrated in vacuo. Thecrude product was purified by preparative HPLC to afford the desiredproduct. (Calc'd for C21H18N4OS3: 74.46, [M+H]+ found: 375).

Preparation of Compounds 152, 194, 195, 196, 197, 198, 199, 201 and 202was done in an analogous manner to Compound 200.

Preparation of Compound 286

To a solution of 5-methyl-2-nitrobenzoic acid (2.0 g, 11.0 mmol) in 100mL of CH₂Cl₂ was added oxalyl chloride (7.0 g, 4.8 mL, 55.2 mmol) and 3drops of DMF. The mixture was stirred at rt for 40 minutes and thesolvents were removed in vacuo. The resulting residue was put under highvacuum, and then dissolved in 10 mL of CH₂Cl₂. This solution was addedto a solution of 2-amino-3-hydroxypyridine (1.21 g, 11.0 mmol) anddiisopropylethylamine (2.1 g, 2.9 mL, 16.6 mmol) in 100 mL of CH₂Cl₂.The reaction was stirred at rt until completion, and was thenpartitioned between CH₂Cl₂ and brine. The organic layer was washed withbrine and dried over MgSO₄. The crude product was purified by silicachromatography, EtOAc/Hexanes, (20:80 to 90:10) to afford the desiredamide product. (Calc'd for C13H11N3O4: 273.25, [M+H]+ found: 274.1).

A slurry of palladium on carbon (10% w/w, 0.12 g) in 1 mL of EtOH wasadded to a solution of the pyridyl amide (1.2 g, 4.4 mmol) in 50 mL ofMeOH. The reaction was placed under a H₂ atmosphere and was stirred atrt overnight. The mixture was purged with nitrogen and then filteredthrough Celite. The volatiles were removed in vacuo to afford thedesired amine. (Calc'd for C13H13N3O2: 243.27, [M+H]+ found: 244).

2-Amino-N-(3-hydroxypyridin-2-yl)-5-methylbenzamide (0.2 g, 0.8 mmol)was combined with 1 mL of PPA, and the mixture was stirred at 150° C.for 2 h. The reaction was quenched with ice and basified with sat.Na2CO3. The resulting yellow solid was filtered, washed with water anddried to afford the desired aniline. (Calc'd for C13H11N3O: 225.25,[M+H]+ found: 226.1).

A microwave vial was charged with4-methyl-2-oxazolo[4,5-b]pyridin-2-yl-phenyl amine (40 mg, 0.2 mmol),3,4-dimethoxybenzoyl chloride (35 mg, 0.2 mmol) and 1 mL of pyridine.The mixture was subjected to microwave irradiation at 160° C. for 10minutes. The resulting precipitate was filtered, washed with MeOH anddried to give the desired amide as a white solid. (Calc'd forC22H19N3O4: 389.41, [M+H]+ found: 390.1).

Preparation of Compound 295

To a solution of methyl 4-(bromomethyl)-3-methoxybenzoate (10.0 g, 38.6mmol) in 200 mL of CH₂Cl₂ was added morpholine (3.36 g, 38.6 mmol) anddiisopropylethylamine (5.98 g, 8.1 mL, 46.3 mmol). The solution wasstirred at rt overnight, and the reaction was then partitioned betweenCH₂Cl₂ and water. The organic extract was washed with brine and driedover MgSO₄. Solvent evaporation afforded the desired product as a stickysolid. (Calc'd for C14H19NO4: 265.31, [M+H]+ found: 266.1).

Methyl 4-(morpholinomethyl)-3-methoxybenzoate (10.4 g, 39.1 mmol) wasdissolved in 150 mL of THF and to this was added a suspension of LiOH(4.68 g, 195.4 mmol) in 75 mL of water. The mixture was stirredovernight at rt, and the solvents were then removed in vacuo. Theresulting solid was suspended in CH₂Cl₂ (200 mL) and MeOH (50 mL). Thismixture was then filtered through Celite, and the solvent was evaporatedfrom the mother liquor to afford the desired acid product. (Calc'd forC13H17NO4: 251.28, [M+H]+ found: 252.1)

The benzoic acid (0.1 g, 0.4 mmol) was suspended in CH₂Cl₂ (5 mL) and tothis was added oxalyl chloride (0.25 g, 2.0 mmol, 0.17 mL) and 2 dropsof DMF. After 1 h at rt, the solvents were removed in vacuo, and theresulting residue was placed under high vacuum for 30 minutes. It wasthen mixed with pyridine (2 mL) and added to a microwave vial chargedwith 3-oxazolo[4,5-b]pyridin-2-yl-phenylamine (0.084 g, 0.4 mmol). Thereaction mixture was subjected to microwave irradiation at 160° C. for10 minutes. The solvents were then removed and the crude material waspurified by silica chromatography (5-10% MeOH/CH₂Cl₂) and prep tlc (5%MeOH/CH₂Cl₂) to afford the desired amide product. (Calc'd forC25H24N4O4: 444.5, [M+H]+ found: 445.1).

Compounds 706, 571, 572, 585, 629, 630, 631, 632, 636, 637, 638, 642 and643 was prepared in an analogous manner to Compound 295.

Preparation of Compound 468

3,5-Dimethyl benzoic acid (0.3 g, 2 mmol) and thionyl chloride (8 mL)were combined under a nitrogen atmosphere and heated to reflux for 3 h.The excess thionyl chloride was evaporated completely in vacuo. To thiswas then added 4-oxazolo[4,5-b]pyridin-2-yl pyridine-2-ylamine (0.318 g,1.5 mmol), dry pyridine (8 mL) and a catalytic amount of DMAP, and themixture was heated to reflux at 110° C. for 6 h. To this water (20 mL)was added and the compound was extracted into CH₂Cl₂ (2×25 mL). Theorganic layer was dried over anhydrous Na₂SO₄, filtered andconcentrated. The crude compound was purified on silica (60-120 mesh)eluted with EtOAc:hexanes (1:5), to obtain the pure amide. (Calc'd forC20H16N4O2: 344.38, [M+H]+ found: 3445.1).

Preparation of Compounds 367, 369, 375, 376, 470, 482, 483, 566, 567,576, 578, 579, 580 and 639 was done in an analogous manner as Compound468.

Preparation of Compound 565

To a solution of 4-oxazolo[4,5-b]pyridin-2-yl pyridine-2-ylamine (0.192g, 0.91 mmol) in dry pyridine (10 mL) was added TMSCl (1 mL 0.75 mmol)at 10-15° C. and the reaction mixture was stirred at room temperaturefor 4 h.

In a separate round bottomed flask, trifluoromethoxy benzoic acid (0.250g, 1.21 mmol) and thionyl chloride (6 mL) were combined under a nitrogenatmosphere and heated to reflux for 3 h. The excess thionyl chloride wasevaporated in vacuo. To this reaction mixture, the above preparedsilylated amine was added at room temperature and the mixture was heatedto reflux for 6 h. Water (20 mL) was added and the compound wasextracted with dichloromethane (2×25 mL). The organic layer was driedover anhydrous Na₂SO₄, filtered and concentrated. The crude compound waspurified on silica (60-120 mesh) eluted with EtOAc:hexanes, to obtainthe pure amide. (Calc'd for C19H11F3N4O3: 400.32, [M+H]+ found: 400.8).

Preparation of Compounds 577, 581, 640, 641 and 668 was done in ananalogous manner to Compound 565.

Preparation of Compound 705

4-Oxazolo[4,5-b]pyridin-2-yl pyridine-2-ylamine (0.20 g, 0.9 mmol), drypyridine (1 mL) and 4-chlorophenylsulphonyl chloride (0.9 g, 0.9 mmol)were mixed in a sealed tube and heated at 220° C. for 2 h. Aftercompletion of the reaction (monitored by TLC), the reaction mixture wascooled to room temperature, water (20 mL) was added, the compound wasextracted into dichloromethane (2×30 mL), and the organic layer wasdried (Na₂SO₄), filtered and concentrated. The crude compound waspurified by silica column chromatography to obtain the pure sulfonamide.(Calc'd for C17H11ClN4O3S: 386.82, [M+H]+ found: 387.0).

Preparation of Compounds 370, 371, 372, 373, 374, 471, 474, 476, 477,478, 479 and 653 was done in an analogous manner to Compound 705.

Preparation of Compound 379

To a stirred solution of 2-oxazolo[4,5-b]pyridin-2-yl-phenylamine (0.2g, 0.94 mmol) in 2 mL of dry pyridine was added 3,5-dimethyl sulfonylchloride (0.193 g, 0.94 mmol) and a catalytic amount of DMAP. Themixture was stirred for 3 h at 140° C. (The progress of the reaction wasmonitored by TLC). The reaction mixture was diluted with water (20 mL),extracted with ethyl acetate (2×25 mL). The combined organic layers weredried over Na₂SO₄ and evaporated in vacuo. The crude compound waspurified by silica chromatography to obtain the correspondingsulfonamide (Calc'd for C20H17N3O3S: 379.44, [M+H]+ found: 380.0).

Preparation of Compounds 377, 378, 380, 381, 382, 383, 384, 486, 487,488, 489, 490, 492, 493 and 494 was done in an analogous manner toCompound 379.

Preparation of Compound 495

To a stirred solution of 2-oxazolo[4,5-b]pyridin-2-yl-phenylamine (0.15g, 0.70 mmol) in 1.5 mL of pyridine was added 4-chlorophenylcholoroformate (0.16 g, 0.84 mmol) followed by a catalytic amount ofDMAP at room temperature. The reaction was stirred for 2 h under anitrogen atmosphere. (The progress of the reaction was monitored byTLC). The reaction mixture was diluted with water (10 mL), extractedwith ethyl acetate (2×25 mL) and the combined organic layers were driedover anhydrous Na₂SO₄, filtered and concentrated in vacuo. The crudecompound was purified by column chromatography to obtain thecorresponding carbamate. (Calc'd for C19H12N3O3Cl: 365.7, [M+H]+ found:366.0).

Preparation of Compounds 496, 497, 499, 500, 501, 502, 568 and 582 wasdone in an analogous manner to Compound 495.

Preparation of Compound 584

Triphosgene (0.72 g, 2.4 mmol) was dissolved in dry dichloromethane andpyridine (1 equiv) was added at −78° C. The mixture was stirred for 10min. To this reaction mixture was added 3,5-dimethyl phenol (0.5 g, 4mmol) in 2 mL of DCM over a period of 10 min. and stirred for 2 h atroom temperature. In another RB flask,2-oxazolo[4,5-b]pyridin-2-yl-phenylamine (0.2 g, 0.9 mmol) was dissolvedin pyridine (20 mL) and to this was added the above prepared phenylchloroformate drop wise over a period of 5 min at room temperature. Acatalytic amount of DMAP was added and the reaction was stirredovernight. After complete disappearance of the starting material(monitored by TLC), the reaction mixture was diluted with water (10 mL)and extracted with ethyl acetate (2×25 mL). The combined organic layerswere dried over anhydrous Na₂SO₄, filtered and concentrated in vacuo.The crude compound was purified by silica column chromatography toobtain the pure compound. (Calc'd for C21H17N3O3: 359.38, [M+H]+ found:360.0).

Preparation of Compounds 569, 570, 583, 655, 656, 657, 669, 670 and 671was done in an analogous manner to Compound 584.

Preparation of Compound 672

To a stirred solution of 4-trifluoromethoxy phenol (0.1 g, 0.56 mmol) indry DMF (0.5 mL), was added N,N-carbonyl diimidazole (0.09 g, 0.56mmol). The reaction was stirred for 30 min at room temperature under anitrogen atmosphere. The reaction mixture was diluted with water (10mL), extracted into ethyl acetate, dried over anhydrous Na₂SO₄, filteredand evaporated in vacuo. This crude compound was dissolved in pyridine(0.5 mL) and was added to a pre-stirred solution of2-oxazolo[4,5-b]pyridin-2-yl-phenylamine (0.075 g, 0.35 mmol) inpyridine (0.5 mL) followed by the addition of a catalytic amount ofDMAP. The mixture was stirred for 2 h at room temperature. Aftercompletion of the reaction (monitored by TLC), the reaction mixture wasdiluted with water (10 mL), extracted with ethyl acetate, dried overanhydrous Na₂SO₄, filtered and concentrated in vacuo. The crude compoundwas purified by column chromatography to afford the pure compound.(Calc'd for C₂₀H₁₂F3N3O4: 415.33, [M+H]+ found: 415.1).

Preparation of Compound 666

To a solution of 4-oxazolo[4,5-b]pyridin-2-yl pyridine-2-ylamine (0.24g, 0.96 mmol) in pyridine (2 mL), trimethylsilyl chloride (0.6 mL, 4.33mmol) was added drop wise at room temperature under a nitrogenatmosphere and the mixture was stirred for 6 h. To this was added4-dimethyl aminophenyl isothiocyanate (0.251 g, 1.41 mmol) and acatalytic amount of dimethylamino pyridine. The reaction was refluxedfor 12 h (progress of the reaction was monitored by TLC). The reactionmixture was diluted with water (50 mL) and the compound was extractedwith DCM (2×50 mL). The combined organic layers were dried (Na₂SO₄) andevaporated in vacuo. The crude compound was purified by silicachromatography to afford the corresponding thiourea. (Calc'd forC20H18N6OS: 390.47, [M+H]+ found: 391.0).

Preparation of Compounds 573, 574, 575, 658, 659, 660, 661, 662, 663,664, 665, 667, 673, 674 and 675 was done in an analogous manner toCompound 666.

Preparation of Compound 634

To a stirred solution of the thiourea (0.040 g, 1.05 mmol) in DMF:H₂O (4mL, 2:2), was added an NH₃ solution (30%, 3 mL). The mixture was stirredfor 30 min at room temperature. To this was added a NaIO₄ solution(0.033 g, in 2 mL water) and the reaction mixture was heated at 80° C.for 12 h. 10% NaOH solution (1 mL) was then added at room temperature.The resulting precipitate was filtered, washed with water (2×10 mL),hexanes (2×10 mL), and dried to obtain the required pure guanidine.(Calc'd for C18H13ClN6O: 364.8, [M+H]+ found: 364.9).

Preparation of Compounds 633 and 635 was done in an analogous manner toCompound 634.

Preparation of Compound 506

The methyl-4-methylpyrrolodine-3-methoxybenzoate was prepared followingthe same procedure as the methyl-4-methylmorpholine-3-methoxybenzoatewas prepared.

To a suspension of 3-oxazolo[4,5-b]pyridin-2-yl-phenylamine (0.10 g, 0.5mmol) in 1 mL of toluene was added trimethylaluminum (2.0 M in toluene,0.2 mL, 0.5 mmol). This mixture was stirred for 1.5 h at rt. To this wasthen added a slurry of methyl-4-methylpyrrolodine-3-methoxybenzoate(0.12 g 0.5 mmol) in 1 mL of toluene. The reaction was stirred at refluxfor 17 h. Upon cooling, the mixture was partitioned between CH₂Cl₂ andbrine. The aqueous layer was extracted with CH₂Cl₂, and the combinedorganic extracts were washed with brine and dried over MgSO₄. The crudeproduct was purified by silica chromatography, 0 to 10% MeOH/CH₂Cl₂ toafford the desired amide. (Calc'd for C25H24N4O3: 428.50, [M+H]+ found:429.1).

Preparation of Compound 525

3.27 g (30 mmol) of 2,3-diaminopyridine, 4.53 g (30 mmol; 1.0 equiv) of2-nitrobenzaldehyde and 1.92 g (60 mmol; 2.0 equiv) of sulfur werethoroughly mixed together and heated to 120° C. The mixture turned intoa black liquid and was stirred for a further 3 h. After cooling to roomtemperature, the solid residue was dissolved in hot ethanol (400 ml) andfiltered. The filtrate was concentrated, and the residue was purified byflash chromatography (gradient hexane/ethyl acetate 1:1 to 0:100) togive the pyridoimidazole product. (Calc'd for C12H8N4O2: 240.2, [M+H]+found: 242).

In a two-necked 100 mL flask, 604 mg (13.8 mmol; 1.1 equiv) of sodiumhydride (55% in paraffin) under nitrogen were washed with anhydroushexane (2×5 ml) and suspended in anhydrous DMF (30 ml). 3.02 g (12.6mmol) of imidazole 22 were added in portions at 0° C., leading to gasevolution and the formation of a deep red-brown solution. The mixturewas stirred for 30 min at 0° C. and for 30 min at room temperature,cooled to 0° C., and 2.45 ml (13.8 mmol; 1.1 equiv)2-(chloromethoxy)ethyltrimethylsilane were added drop wise. Afterstirring for 5.5 h at room temperature the orange-brown suspension wasadded to a mixture of saturated aq. Na₂CO₃-solution (150 ml), water (300ml), ethyl acetate (200 ml), and brine (50 ml). The aqueous layer wasextracted with ethyl acetate (4×100 ml); and the combined organic layerwas washed with a mixture of brine and water (1:3; 2×100 ml) and brine(50 ml), dried (Na₂SO₄), and concentrated. The residue was purified byflash chromatography (gradient hexane/ethyl acetate 1:2 to 0:100),affording three isomers of the product: 1.90 g (5.13 mmol, 41%), 1.31 g(3.55 mmol; 28%), and 895 mg (2.41 mmol; 19%). (Calc'd for C18H22N4O3Si:370.1, [M+H]+ found: 371.2).

The 2-(2-nitrophenyl)imidazopyridine 1.87 g (5.05 mmol) was dissolved inethyl acetate (27 ml) and methanol (18 ml) in the presence of 10%palladium on charcoal (190 mg) at ambient pressure and temperature for 4h. Purification of the crude product by flash chromatography(hexane/ethyl acetate 1:1) gave the desired aniline as a yellow-greenoil. (Calc'd for C18H224N4OSi: 340.1, [M+H]+ found: 341.2).

The reaction of2-[3-(2-trimethylsilanyl-ethoxymethyl)-3H-imidazo[4,5-b]pyridin-2-yl]phenylamine(85.1 mg; 0.25 mmol) with 3,4-dimethoxybenzoylchloride (60.2 mg; 1.2equiv) and 38 μL (1.1 equiv) of NEt3 in dry CH₂Cl₂ (2 ml), followed byflash chromatography (hexane/ethyl acetate 2:1), afforded the desiredamide as a colorless, highly viscous resin, which solidified uponaddition of ethanol. (Calc'd for: C27H32N4O4Si: 504.2, [M+H]+ found:505).

Preparation of Compounds 521, 522, 523, 524, 526, 527, 528, 529, 530,531, 532, 533, 534, 535, 536, 537, 538, 539 and 540 was done in ananalogous manner to Compound 525.

Preparation of Compound 253

In a typical run, 2-(2-Aminophenyl)indole (104 mg, 0.5 mmol) in 4 mLpyridine was stirred with 2,4-dimethoxybenzoyl chloride (100 mg, 0.5mmol). The reaction was stirred at room temperature overnight. Reactioncompletion was confirmed by LC-MS, and 15 mL of water was charged. Theresulting suspension was stirred and sonicated until white precipitateswere obtained. The white solid was collected by filtration, washed withwater and air dried. The product was purified by flash chromatography onsilica gel using CH₂Cl₂ as eluent (0 to 10% methanol gradient).TLC/HPLC/LC-Mass suggested that it was the clean product (MS,M⁺+H=373.1).

Compounds 254, 255 and 256 were prepared in an analogous manner toCompound 253, using the appropriate acid chlorides.

Preparation of Compound 262

In a typical run, 4-(1H-Benzoimidazol-2-yl)-phenylamine (83 mg, 0.4mmol) in 3 mL pyridine was added to 2,4-dimethoxybenzoyl chloride (80mg, 0.4 mmol). The reaction was stirred overnight at room temperature.Reaction completion was confirmed by LC-MS, and 15 mL of water wascharged. The resulting suspension, upon stirring for five hours, wasfiltered to collect an off white solid, which was washed with water anddried under reduced pressure. The product was purified by flashchromatography on silica gel using CH₂Cl₂ as eluent (0 to 10% methanolgradient). TLC/HPLC/LC-Mass suggested that it was the clean product (MS,M⁺+H=374.1).

Compounds 263, 264 and 294 were prepared in an analogous manner toCompound 262, using the appropriate acid chlorides, and purified eitherby recrystallization from acetonitrile or normal phase chromatographyusing a 9:1 mixture of CH₂Cl₂ to MeOH as eluent.

Preparation of Compound 332

In a typical run to prepare Compound 332, in a vial was dissolved2-Pyridin-3-yl-phenylamine (400 umol, 68 mg) in 2 ml of pyridine.3,4-dimethoxybenzoyl chloride (400 umol, 80 mg) was added with stirring.The solution was stirred overnight at room temperature. 20 ml H₂O wasthen added and the resulting white emulsion was stirred overnight withintermittent sonication until white precipitate was obtained in a clearsolution. The solid was filtered, and air dried. Purification was doneon silica gel with in CH₂Cl₂ (methanol gradient 0 to 10%), concentratedto dryness to obtain the product as a white solid. (MS, M⁺+H=335.1).

Compounds 331 and 333 were prepared in an analogous manner to Compound332, using the appropriate acid chlorides. In the cases of Compounds 331and 333, the crude product was isolated by extraction into CH₂Cl₂.

Preparation of Compound 334

In a typical run to prepare Compound 332, in a vial was dissolved2-Pyridin-3-yl-phenylamine (400 umol, 70 mg) in 2 ml of pyridine.2,4-dimethoxybenzoyl chloride (400 umol, 80 mg) was added with stirring.The solution was stirred overnight at room temperature. 20 ml H₂O wasthen added and the resulting white emulsion was stirred overnight withintermittent sonication until white precipitate was obtained in a clearsolution. The solid was filtered, and air dried. Purification wasperformed on silica gel with CH₂Cl₂ (methanol gradient 0 to 10%),concentrated to dryness to obtain the product as a white solid. (MS,M⁺+H=340.1).

Compounds 335 and 336 were prepared in an analogous manner to Compound332, using the appropriate acid chlorides. In the case of Compound 335,the crude product was isolated by extraction into CH₂Cl₂.

Preparation of Compound 413

In a typical run to prepare Compound 413, in a vial was dissolved2-Pyridin-3-yl-phenylamine (300 umol, 66 mg) in 1 ml of pyridine.2,4-dimethoxybenzoyl chloride (300 umol, 60 mg) was added with stirring.The solution was stirred overnight at room temperature, concentrated todryness, and chased with pentane (2×5 ml) to obtain the crude product asan orange solid. The crude was purified by silica gel chromatography (0%to 70% Ethyl Acetate in Pentane) to obtain the desired product (77 mg).(MS, M⁺+H=385.1)

Compounds 414, 415 and 416 were prepared in an analogous manner toCompound 413, using the appropriate acid chlorides.

Preparation of 2-(2-Nitro-phenyl)-imidazo[1,2-a]pyridine

2-Bromo-2′-nitroacetophenone (7.65 g, 31.3 mmol) and 2-aminopyridine(2.95 g, 31.3 mmol) were dissolved in acetone (50 ml) and brought toreflux. After 10 minutes a light yellow/white precipitate forms inaddition to strong boiling. Refluxing was continued for 3 hours andcooled to room temperature. The volume was reduced ½ by evaporation andthe intermediate solids were collected by filtration and washed withacetone (20 ml) and air dried (7.26 g of intermediate). The intermediate(7.20 g) was dissolved in MeOH (100 ml) with HBr (4 drops, catalytic)and brought to reflux. The reaction was monitored by TLC (10% MeOH inCH₂Cl₂). After 70 minutes, the reaction mixture was cooled to roomtemperature, adjusted to pH=12 with 1M NaOH, and concentrated to removethe methanol. The yellow product was collected by filtration, washedwith water and air dried. 5.54 g (74%) of2-(2-Nitro-phenyl)-imidazo[1,2-a]pyridine was obtained as a yellowcrystalline solid (MS, M⁺+H=240.1).

Preparation of 2-Imidazo[1,2-a]pyridin-2-yl-phenylamine

2-Imidazo[1,2-a]pyridin-2-yl-phenylamine was prepared with two reductivemethods.

1) Catalytic hydrogenation. 2-(2-Nitro-phenyl)-imidazo[1,2-a]pyridine(250 mg, 1.04 mmol) was dissolved in THF (15 mL). The flask was charged20 mg of 10% Palladium on Carbon, flushed with nitrogen and stirred overH₂ balloon (1 atm.) O/W. The reaction was monitored by either HPLC orTLC (5% MeOH in CH₂Cl₂). The reaction mixture was filtered over a bed ofCelite to remove the catalyst, the bed was washed with THF (2×10 ml) andthe combined organics were concentrated to dryness to obtain an amberoil. The white solid product was obtained by careful roto-evaporation ofa 50% Aqueous ethanol solution, and collected by filtration (MS,M⁺+H=210.1).

2) Sulfide reduction. Into a round bottom flask was charged2-(2-Nitro-phenyl)-imidazo[1,2-a]pyridine (250 mg, 1.04 mmol), Sodiumhydrogensulfide (351 mg, 6.24 mmol), methanol (6 ml) and water (2 ml).The reaction mixture was refluxed overnight. TLC indicated that thereaction was complete (5% MeOH in CH₂Cl₂). The mixture was cooled toroom temperature, concentrated to dryness and to the white/yellow saltswas added water (1 mL), CH₂Cl₂ (10 mL) and MeOH (1 mL). The layers wereseparated and the aqueous layer was back extracted with CH₂Cl₂ (2×10ml). The combined organic layers were dried over Na₂SO₄, andconcentrated to dryness to obtain the product as a tan solid (MS,M⁺+H=210.1).

Preparation of Compound 265

In a typical run to prepare Compound 265, in a vial was dissolved2-Imidazo[1,2-a]pyridin-2-yl-phenylamine (400 umol, 84 mg) in 3 ml ofpyridine. 2,4-dimethoxybenzoyl chloride (400 umol, 80 mg) was added withstirring. The solution was stirred overnight at room temperature. 15 mlH₂O was then added and the resulting white emulsion was stirred for 5hours with intermittent sonication until a white precipitate wasobtained in a clear solution. The solid was filtered, and air dried.Purification was performed on silica gel with CH₂Cl₂ as eluent (methanolgradient 0 to 10%), concentrated to dryness and triturated with Pentaneto obtain the product as an off white solid (MS, M⁺+H=374.1).

Compounds 266, 267 and 268 were prepared in an analogous manner toCompound 265, using the appropriate acid chlorides.

Preparation of 2-(2-Nitro-phenyl)-imidazo[1,2-a]pyridine-3-carbaldehyde

In a dry flask charged with DMF (2.7 g, 37 mmol) and cooled to 0° C. wasadded slowly POCl₃ over 5 minutes. The solution was stirred for 10 minat 0° C. was warmed to room temperature over 1 hour. To the deep redcolor solution recooled to 0° C. was added2-(2-Nitro-phenyl)-imidazo[1,2-a]pyridine (1.0 g, 4.18 mmol) in DMF (8ml). The reaction was stirred for 5.5 hours; LC-MS indicated someproduct formation. A second addition of the Vilsmeier complex (from 2.7g of DMF and 1.47 g POCl₃) was charged and the reaction stirredovernight, then heated to 50° C. for 3 hours until reaction completion.The reaction was cooled to room temperature, poured onto ice, adjustedto pH=7 with 1N NaOH. The product was extracted into CH₂Cl₂ (3×50 ml),washed with brine, dried over Na₂SO₄ and concentrated to obtain theproduct as a white solid (1.03 g, 92% yield). (MS, M⁺+H=268.0)

Preparation of Compound 316

To 2-(2-Nitro-phenyl)-imidazo[1,2-a]pyridine (700 mg, 2.62 mmol)suspended in methanol (30 ml) cooled to 0° C. was charged a solution ofNaBH₄ (99 mg, 2.62 mmol) in methanol (2 ml). After stirring at 0° C. for15 min, the solution was warmed to room temperature and stirred for 1.5hours. The pH was adjusted to 6 with 4 N HCl, and the solutionconcentrated to remove the methanol. The yellow solid was collected byfiltration, washed with water and dried thoroughly to obtain the productas a yellow solid (539 mg, 76% yield). (MS, M⁺+H=270.1)

Preparation of 3-Chloromethyl-2-(2-nitro-phenyl)-imidazo[1,2-a]pyridine

To [2-(2-Nitro-phenyl)-imidazo[1,2-a]pyridin-3-yl]-methanol (500 mg,1.86 mmol) suspended in CH₂Cl₂ (25 ml) was added dropwise thionylchloride (662 mg, 3 eq). The reaction was stirred at room temperaturefor 3.5 hours, concentrated to dryness, chased with CH₂Cl₂ and driedunder reduced pressure to obtain the product as a white solid inquantitative yield. (MS, M⁺+H=270.1, reacts with water diluent.

Preparation of2-(3-Dimethylaminomethyl-imidazo[1,2-a]pyridin-2-yl)-phenylamine

To 3-Chloromethyl-2-(2-nitro-phenyl)-imidazo[1,2-a]pyridine (324 mg, 1.0mmol) suspended in CH₂Cl₂ (20 ml) cooled to 0° C. was added dropwisetriethylamine (303 mg, 3 eq), followed by 2 M dimethylamine in THF (8mmol, 4 ml) in two portions over a 5 hour period. The reaction mixturewas concentrated to dryness, dissolved in CH₂Cl₂ (20 ml) washed withwater, brine, dried over Na₂SO₄, concentrated and purified bychromatography using a 9:1 mixture of CH₂Cl₂/MeOH. The fractions wereconcentrated, dissolved in THF (15 ml), and stirred over H₂ balloon with10% Pd/C (10 mg) overnight. The reaction mixture was filtered throughCelite and concentrated to obtain the product as a yellow solid (164mg). (MS, M⁺+H=267.1)

Preparation of Compound 350

In a typical run to prepare Compound 350,2-(3-Dimethylaminomethyl-imidazo[1,2-a]pyridin-2-yl)-phenylamine (200umol, 53 mg) in 2 ml of pyridine was stirred with 3,4-dimethoxybenzoylchloride (200 umol, 40 mg) at room temperature overnight. The reactionmixture was quenched by the addition of 10 ml H₂O, concentrated andpurified by chromatography using a 90:9:1 mixture ofCH₂Cl₂/Methanol/Triethylamine. Trituration with pentane gave the productas a tan solid. Higher purity can be achieved with additional silica gelcolumn chromatography (100% EtOAc) and/or prep TLC (5% MeOH in CH₂Cl₂w/7M NH₃). (MS, M⁺+H=431.1)

Compound 351 was prepared in an analogous manner to Compound 350, usingthe appropriate acid chlorides.

Preparation of4-[2-(2-Amino-phenyl)-imidazo[1,2-a]pyridin-3-ylmethyl]-piperazine-1-carboxylicacid tert-butyl ester

To 3-Chloromethyl-2-(2-nitro-phenyl)-imidazo[1,2-a]pyridine (298 mg,0.92 mmol) suspended in CH₂Cl₂ (15 ml) cooled to 0° C. was addeddropwise triethylamine (279 mg, 3 eq), followed by 1-Boc-piperazine (1.3eq, 221 mg) in two portions over a 24 hour period. The reaction mixturewas concentrated to dryness and purified by chromatography using a 9:1mixture of CH₂Cl₂ to MeOH. The fractions were concentrated, dissolved inEthanol (20 ml), and stirred over H₂ atmosphere with 10% Pd/C (10 mg)for 48 hours. The reaction mixture was filtered through Celite andconcentrated, and dried under high vacuum to obtain 250 mg of thedesired product. (MS, M⁺+H=408.2)

Preparation of Compound 359

In a typical run to prepare Compound 359,4-[2-(2-Amino-phenyl)-imidazo[1,2-a]pyridin-3-ylmethyl]-piperazine-1-carboxylicacid tert-butyl ester (150 umol, 61 mg) in 2 ml of pyridine was stirredwith 2,4-dimethoxybenzoyl chloride (150 umol, 30 mg) for 3 hours at roomtemperature. The reaction was quenched with 1 ml H2O, concentrated andpurified by silica gel chromatography (0 to 5% MeOH in CH₂Cl₂). Thefractions were concentrated, treated with TFA/CH₂Cl₂ for 5 hours,concentrated to dryness, chased with CH₂Cl₂ (2×10 mL), pumped under highvacuum and triturated with 1:1 Pentane/Ether to obtain the Bis TFA saltas a white solid, (46 mg). (MS, M⁺+H=472.1)

Compounds 362 and 364 were prepared in an analogous manner to Compound359, using the appropriate acid chlorides.

Preparation of 2-(2-Nitro-phenyl)-imidazo[1,2-a]pyrimidine

2-Bromo-2′-nitroacetophenone (5.0 g, 20.4 mmol) and 2-aminopyrimidine(1.94 g, 20.4 mmol) were dissolved in acetone (50 ml) and brought toreflux. After 60 minutes a light white precipitate developed. Refluxingwas continued for 5.5 hours and cooled to room temperature. The volumewas reduced ½ by evaporation and the intermediate solids were collectedby filtration, washed with acetone (20 ml) and air dried (2.84 g ofintermediate). A second crop was collected by concentration of thecombined mother liquors, and refluxing the residue in fresh acetone (20ml) for 1.5 hours. Upon cooling, and reducing the volume by ½ byevaporation, 3.11 g of solids were collected. The combined solidintermediate (5.95 g) was dissolved in MeOH (70 ml) with 5 drops ofconc. HBr and refluxed for 2 hours. After cooling the volume was reducedby ½, water (40 ml) was added solution was made basic (pH=10) with 1NNaOH. Roto-evaporation to remove methanol resulted in the productcrystallizing. The product was filtered, washed with water and air driedto obtain 5.03 g of the desired product. (MS, M⁺+H=241.0)

Preparation of 2-Imidazo[1,2-a]pyrimidin-2-yl-phenylamine

2-(2-Nitro-phenyl)-imidazo[1,2-a]pyrimidine (2 g, 8.33 mmol) and NaHS(2.8 g, 6 eq) were refluxed in 25% aqueous methanol (40 ml) waterovernight. The reaction mixture was cooled to room temperature, andconcentrated to dryness. Methanol (10 ml) and water (30 ml) were added,and the product was extracted with CH₂Cl₂ (3×100 ml). The combinedorganic layers were concentrated to dryness, chased with 10% MeOH:CH₂Cl₂and dried under high vacuum at 45° C. to obtain the product as an orangesolid (1.34 g, 76% yield). (MS, M⁺+H=211.1)

Preparation of Compound 437

In a typical run to prepare Compound 437,2-Imidazo[1,2-a]pyrimidin-2-yl-phenylamine (200 umol, 42 mg) in 2 ml ofpyridine was stirred with 2,4-dimethoxybenzoyl chloride (200 umol, 40mg) overnight at room temperature. The following morning the reactionmixture was concentrated to dryness, chased with CH₂Cl₂ and Pentane.Purification was performed on silica gel with a 30% to 70% EtOAcgradient in Pentane. (MS, M⁺+H=375.1)

Compounds 438, 439, 504 and 505 were prepared in an analogous manner toCompound 437, using the appropriate acid chlorides.

Preparation of2-(5,6,7,8-Tetrahydro-Imidazo[1,2-a]pyridin-2-yl)-phenylamine

In a dry flask was charged 2-(2-Nitro-phenyl)-imidazo[1,2-a]pyridine(239 mg, 1 mmol), 10% Pd/C (15 mg), Ethanol (25 ml), water (1.5 ml) and4M HCl (0.5 ml). The atmosphere was flushed with nitrogen and stirredover 1 atm. hydrogen over 5 days with an additional charge of Pd/Ccatalyst (15 mg) and 4 N HCl (4.5 ml) until reaction completion wasobtained. The reaction mixture was filtered through Celite, andneutralized to pH=6 with a saturated NaHCO₃ solution. The solution wasroto-evaporated to remove the ethanol, the aqueous layer was extractedwith CH₂Cl₂ (2×5 ml), and the combined organic layer dried over Na₂SO₄and concentrated to obtain the product as an oil in quantitative yield(205 mg). (MS, M⁺+H=214.1)

Preparation of Compound 545

In a vial was added2-(5,6,7,8-Tetrahydro-imidazo[1,2-a]pyridin-2-yl)-phenylamine (200 umol,42 mg) and triethylamine (3.5 eq, 100 ul) in anhydrous CH₂Cl₂ (3 ml)followed by 3,4-dimethoxybenzoyl chloride (200 umol, 40 mg) withstirring. The reaction was stirred overnight at room temperature,concentrated to dryness, and purified on silica gel with a 0% to 100%EtOAc gradient in Pentane to obtain the product as a white solid (30 mg)after trituration with pentane. (MS, M⁺+H=378.1)

Compounds 546, 547 and 548 were prepared in an analogous manner toCompound 545, using the appropriate acid chlorides.

Preparation of Compound 541

In a vial charged with 3-Methoxy-4-morpholin-4-ylmethyl-benzoic acid(125 mg, 500 umol) dissolved in anhydrous CH₂Cl₂ (5 mL) was added oxalylchloride (0.5 mL, 11 eq) and 1 drop of DMF. After stirring for 3 hoursthe reaction was concentrated to dryness to obtain the acid chloride. Tothe acid chloride was added a solution of2-Imidazo[1,2-a]pyridin-2-yl-phenylamine (500 mmol, 104 mg) in anhydrousPyridine (5 ml). After stirring for 2 hours the reaction mixture wasconcentrated to dryness, suspended in EtOAc, and washed with 50%saturated NaHCO₃ solution. The aqueous layer was back extracted withEtOAc and the combined organic layers were dried over Na₂SO₄ andconcentrated to obtain the crude yellow solid. Column chromatography (40g silica, 0% to 5% MeOH gradient in CH₂Cl₂), concentration andtriturating in methanol gave the product as a white solid (57 mg). (MS,M⁺+H=443.1)

Compounds 542 was prepared in an analogous manner to Compound 541, using2-Imidazo[1,2-a]pyrimidin-2-yl-phenylamine.

Preparation of2-(3-Pyrrolidin-1-ylmethyl-imidazo[2,1-b]thiazol-6-yl)-phenylamine

[6-(2-Nitro-phenyl)-imidazo[2,1-b]thiazol-3-yl]-methanol (110 mg, 0.4mmol) in CH₂Cl₂ (5 ml) with triethylamine (56 ul, 1 eq) was cooled to 0°C. Methylsulfonyl chloride (31 ul, 1 eq) was added dropwise, stirred at0° C. for 10 min, warmed to room temperature, and stirred for 15 min.The reaction was quenched by the addition of brine, and the mesylate wasextracted with CH₂Cl₂, dried over Na₂SO₄ and concentrated. The residuewas dissolved in acetonitrile (3 ml), and added triethylamine (31 ul, 1eq), followed by pyrrolidine (66 ul, 2 eq). The reaction mixture wasstirred for 2 hours, concentrated and chased with pentane. Columnchromatography in CH₂Cl₂ (0 to 4% MeOH gradient) gave6-(2-Nitro-phenyl)-3-pyrrolidin-1-ylmethyl-imidazo[2,1-b]thiazole. Thismaterial was dissolved in methanol (16 ml) and a solution of sodiumhydrogen sulfide (112 mg, 5 eq) in water (4 ml) was added. The reactionmixture was refluxed for 2 days with additional charges of NaHS (2×112mg). The reaction was concentrated to remove the methanol, and theaqueous solution was extracted with CH₂Cl₂ (3×40 ml). The organic layerwas dried over Na₂SO₄, and concentrated to obtain the product as ayellow film, 109 mg. (MS, M⁺+H=299.1)

Preparation of Compound 620

2-(3-Pyrrolidin-1-ylmethyl-imidazo[2,1-b]thiazol-6-yl)-phenylamine (200umol) in 2 ml of pyridine was stirred with 3,4,5-trimethoxybenzoylchloride (200 umol, 46 mg). The solution was stirred for 3 hours,concentrated to dryness, chased with Methanol and purified by prep-HPLC.The fractions were lyophilized to obtain 36 mg of the product as a TFAsalt. (MS, M⁺+H=493.1.)

Compound 619 was prepared in an analogous manner to Compound 620, usingthe appropriate acid chlorides.

Preparation of 6-(2-Nitro-phenyl)-imidazo[2,1-b]thiazole-3-carboxylicacid ethyl ester

In a typical run, ethyl (2-Amino-imidazol-4-yl)-acetic acid methyl ester(1.0 g, 5.8 mmol) was mixed with 30 mL of methyl ethyl ketone along with2-bromo-2′-nitroacetophenone (1.42 g, 5.8 mmol). The reaction mixturewas refluxed for 1 hour and stirred at 90° C. overnight. It was thencooled to room temperature and concentrated to a red oil. Efforts toprecipitate the product by dissolution in methanol and adding waterresulted in an emulsion. Methanol was removed by roto-evaporation andthe aqueous emulsion was charged to a separatory funnel. The pH wasadjusted to 9 with NaHCO₃, and the mixture was extracted with CH₂Cl₂(3×50 ml). The combined organic layer was dried over Na₂SO₄,concentrated to a red oil and purified on silica gel chromatography(CH₂Cl₂ with 0 to 5% MeOH gradient). The product was obtained as a redsolid (0.59 g, 32% yield). (MS, M⁺+H=318.0.)

Preparation of [6-(2-Nitro-phenyl)-imidazo[2,1-b]thiazol-3-yl]-aceticacid

6-(2-Nitro-phenyl)-imidazo[2,1-b]thiazole-3-carboxylic acid ethyl ester(466 mg, 1.47 mmol) in 2:1 THF/Water was combined with 4 eq of NaOH (234mg). The reaction mixture was heated to 50° C. for 3 hours. The reactionmixture was concentrated to dryness, the residue dissolved in water (20ml), washed with CH₂Cl₂, and the aqueous layer was adjusted to pH=3 with4N HCl. The solids were collected by filtration, washed with water andair dried to obtain the acid product was a brown solid (442 mg, 99%yield) (MS, M⁺+H=304.0)

Preparation of Compound 649

In a vial [6-(2-Nitro-phenyl)-imidazo[2,1-b]thiazol-3-yl]-acetic acid(30 mg, 100 uMol), N-methylpiperzine (10 mg, 1.0 eq), andN,N-Diisopropylethylamine (52 uL, 3.0 eq) were dissolved in CH₂Cl₂. HOAT(16 mg, 1.2 eq) was added to the reaction mixture followed by EDCI (29mg, 1.5 eq). The reaction was stirred overnight. After adding 50%saturated NaHCO₃ (2 ml) and extracting with CH₂Cl₂ (3×3 mL), the organiclayer was dried over Na₂SO₄, and concentrated. Trituration with Pentanegave the amide product as a brown solid.

The amide from above was dissolved in methanol (4 ml) with NaHS (34 mg,6 eq) and microwave heated at 150° C. for 30 min. MgSO₄ was charged tothe reaction mixture, and upon filtering, concentration, and chasingwith CH₂Cl₂ (2×) the desired aniline was obtained as a red film.

The aniline from above was dissolved in pyridine (2 ml) and2-Quinoxaloyl chloride (38 mg, 2.0 eq) was charged as a solid. Afterstirring overnight, the reaction was concentrated to dryness andpurified on prep-HPLC to obtain the title compound as an orange solid(32.2 mg, 44% yield over 3 steps). (MS, M⁺+H=512.2)

Preparation of Compound 650

To a vial was added4-[6-(2-Amino-phenyl)-imidazo[2,1-b]thiazol-3-ylmethyl]-piperazine-1-carboxylicacid tert-butyl ester (82 mg, 0.2 mmol), Triethylamine (56 ul, 2 eq) andanhydrous CH₂Cl₂ (3 ml). 2-quinoxaloyl chloride (40 mg, 1.0 eq) wasadded as a solid. The reaction mixture was stirred for 18 hours,concentrated and chased with CH₂Cl₂. Purification on silica gel with aCH₂Cl₂ eluent (with 95:4:1 CH₂Cl₂:MeOH:Et₃N gradient) gave Compound 650as a yellow solid. (MS, M⁺+H=570.2)

Preparation of Compound 651

Compound 650 (105 mg, 0.185 mmol) was treated with 30% TFA in CH₂Cl₂ (4ml) for 2 hours, chased with CH₂Cl₂ (3×) and Ether (3×), to obtain crudeCompound 441. Half of the material (92 umol) was dissolved in CH₂Cl₂ (5ml), along with Et₃N (70 ul) and cooled to 0° C. Acetic anhydride (10ul, 1 eq) was added and the reaction mixture was warmed to roomtemperature over 1 hour. The reaction was quenched by the addition ofmethanol and water, and concentrated to dryness. Purification on reversephase prep HPLC and lyophilization gave Compound 651 as a TFA salt. (MS,M⁺+H=512.2)

Preparation of 2-[6-(2-Nitro-phenyl)-imidazo[2,1-b]thiazol-3-yl]-ethanol

[6-(2-Nitro-phenyl)-imidazo[2,1-b]thiazol-3-yl]-acetic acid (300 mg, 1.0mmol) was suspended in THF (20 ml), and stirred with NMM (110 ul, 1 eq)at room temperature for 1 hour. The reaction mixture was cooled to 0° C.and isobutylchloroformate (131 ul, 1 eq) was charged and the reactionmixture was stirred at 0° C. for 2 hours at which time the mixedanhydride formation was complete. A mixture of NaBH₄ (38 mg) in water (5ml) was added dropwise at 0° C. and warmed to room temperature andstirred for 1 hour. The reaction did not proceed to completion with 1eq.; therefore the NaBH₄ addition was repeated with 3 eq of NaBH₄. Thereaction was not complete after 1 hour, therefore the reaction mixturewas concentrated to dryness, charged with fresh THF, followed by NaBH₄(1 eq) and stirred overnight. LC-MS indicated the reaction was complete,therefore the mixture was concentrated to dryness, and CH₂Cl₂ (50 ml)and water (20 ml) were added. The layers were split, and the aqueouslayer was extracted with CH₂Cl₂ (3×50 ml), and the combined organiclayers were dried over Na₂SO₄ and concentrated to dryness. The productwas purified on silica gel (Pentane with 15% to 100% EtOAc gradient),concentrated and lyophilized from CH₃CN:H₂O (160 mg, 55% yield). (MS,M⁺+H=290.0)

Preparation of4-{2-[6-(2-Nitro-phenyl)-imidazo[2,1-b]thiazol-3-yl]-ethyl}-piperazine-1-carboxylicacid tert-butyl ester

2-[6-(2-Nitro-phenyl)-imidazo[2,1-b]thiazol-3-yl]-ethanol (40 mg, 0.14mmol) was dissolved in anhydrous CH₂Cl₂ and cooled to 0° C.Triethylamine (19 ul, 1 eq) was added followed by Methanesulfonylchloride (11 ul, 1 eq). The reaction mixture was warmed to roomtemperature and stirred for 30 min. LC-MS indicated the reaction wasincomplete, therefore the addition of Triethylamine (19 ul, 1 eq) andmethanesulfonyl chloride (11 ul, 1 eq) was repeated. The reactionmixture was quenched with the addition of 2 ml of brine, extracted withCH₂Cl₂ (2×2 ml), dried over Na₂SO₄ and concentrated to obtain themesylate as a yellow film.

The mesylate was dissolved in anhydrous acetonitrile (2 ml),Triethylamine (38 ul, 2 eq) and stirred with N-Boc-piperazine (26 mg, 2eq) overnight. The reaction mixture was still exclusively the mesylate.The reaction mixture was charged with sodium iodide (41 mg) and stirredfor 6 days and purified by reverse phase prep HPLC. The fractions weremade alkaline with NaHCO₃ (sat), concentrated to remove CH₃CN and theaqueous layer was extracted with CH₂Cl₂. The organic layer was dried andconcentrated to obtain the product as a yellow film. (MS, M⁺+H=458.2)

Preparation of Compound 680

In a microwave tube was added4-{2-[6-(2-Nitro-phenyl)-imidazo[2,1-b]thiazol-3-yl]-ethyl}-piperazine-1-carboxylicacid tert-butyl ester (24 mg, 0.05 mmol), NaHS (30 mg, 10 eq) and 5 mlof methanol. The reaction mixture was microwave heated at 150° C. for 30min. The reaction proceeded, but was not complete. An additional 30 mgof NaHS was charged, and the reaction mixture was microwave heated at150° C. for 30 minutes. Again, charged 15 mg of NaHS and microwaveheated at 160° C. for 20 minutes. The solids were removed by filtration;the solution was dried over MgSO₄, and concentrated to obtain the amineintermediate. This amine (0.05 mmol) was mixed with pyridine (3 ml),with 2-quinoxaloyl chloride (20 mg, 2 eq) and microwave heated at 160°C. for 10 min. The reaction was partially complete, therefore aftercharging another two equivalents of 2-quinoxaloyl chloride (20 mg) thereaction was microwave heated for 20 minutes at 160° C. The reactionmixture was concentrated to dryness, and purified on silica gelchromatography (CH₂Cl₂ eluent, 0 to 5% MeOH gradient). The residue wastreated with 25% TFA/CH₂Cl₂ for 3 hours, concentrated and purified onreverse phase prep HPLC. (MS, M⁺+H=484.2).

Preparation of 3-Chloromethyl-6-(2-nitro-phenyl)-imidazo[2,1-b]thiazole

To a stirred solution of[6-(2-Nitro-phenyl)-imidazo[2,1-b]thiazol-3-yl]-methanol (1.0 g, 3.63mmol) in anhydrous dichloromethane (15 ml) was slowly added thionylchloride (2 ml, 7.5 eq). The solution turned homogenous, followed bydevelopment of a yellow precipitate. After 5 minutes, a catalytic amountof DMF (1 drop) was added and the mixture was stirred for 1 hour,concentrated to dryness, chased with CH₂Cl₂ (2×) then Ether (1×), anddried under reduced pressure. 1.53 g of yellow solid was obtained, andassumed to be quantitative yield. (MS, M⁺+H=294.0)

Preparation of Compound 700

Displacement: 3-Chloromethyl-6-(2-nitro-phenyl)-imidazo[2,1-b]thiazole(126 mg, 0.300 mmol) in 2 ml of 1-methyl-piperazine was microwave heatedat 110° C. for 30 minutes. The reaction mixture was concentrated todryness, and chased with methanol to obtain crude3-(4-Methyl-piperazin-1-ylmethyl)-6-(2-nitro-phenyl)-imidazo[2,1-b]thiazole.

Nitro reduction: The above residue was dissolved in ethanol (20 ml), and10% Palladium on carbon was added with stirring. The atmosphere wasevacuated and backfilled with Nitrogen (3×) and stirred over H₂ balloon(1 atm.) for 18 hours. The reaction mixture was filtered through Celite,concentrated to dryness, and chased with CH₂Cl₂ and pentane to obtainthe amine as a red oil.

Amide Formation: The amine from above was dissolved in pyridine (3 ml),added to a microwave tube containing 2-quinoxalyl chloride (64 mg, 1.1eq) and microwave heated for 30 minutes at 160° C. Reaction was only 50%complete, therefore another portion of 2-quinoxalyl chloride was chargedand heating was continued for 30 minutes at 160° C. The reaction mixturewas concentrated to dryness and purified by reverse phase prep-HPLC.(MS, M⁺+H=512.2)

Compounds 714, 715, 716, and 717 were prepared in an analogous manner toCompound 700, using the appropriate amines. (Boc protecting groups wereremoved by treatment with 25% TFA in CH₂Cl₂ for 3 hours, prior topurification.)

Preparation of Compound 718

6-(2-Nitro-phenyl)-imidazo[2,1-b]thiazole-3-carboxylic acid ethyl ester(0.342 g, 1 mmol) was dissolved in 3:1 Ethanol:THF (80 ml). To thereaction mixture was added 10% Pd/C (30 mg) and the reaction mixture wasstirred over H₂ balloon (1 atm) for 7 days with periodic charges ofadditional catalyst. The reaction mixture was filtered through Celite,concentrated to dryness and chased with pentane to obtain the aniline asan orange solid, 289 mg.

A portion of the aniline from above (56 mg, 200 umol) was dissolved inpyridine (4 ml) and stirred with 2-quinoxalyl chloride (46 mg 1.2 eq)overnight at room temperature. The reaction mixture was quenched withethanol, concentrated to dryness, and chased with CH₂Cl₂/pentane. Theresidue was dissolved in CH₂Cl₂ and washed with 50% saturated aqueousNaHCO₃, dried over Na₂SO₄. The product was purified on silica gel(CH₂Cl₂ with 0 to 5% methanol gradient). (MS, M⁺+H=444.1)

Compound 720 was prepared in an identical manner to Compound 718, using[6-(2-Nitro-phenyl)-imidazo[2,1-b]thiazol-3-yl]-acetic acid methyl esteras the starting material.

Preparation of Compound 719

6-{2-[(Quinoxaline-2-carbonyl)-amino]-phenyl}-imidazo[2,1-b]thiazole-3-carboxylicacid ethyl ester was dissolved in 1:10 THF:Methanol (33 ml) and stirredwith 1 M aqueous NaOH (4 ml) overnight. The reaction was complete byLC-MS. The reaction mixture was concentrated to remove the organics, andcharged with water (20 ml). The alkaline (pH=13) aqueous layer waswashed with CH₂Cl₂ (2×20 ml). The aqueous layer was acidified (pH=2)with 4 M HCl, and extracted with CH₂Cl₂ (3×20 ml). The combined organiclayers were dried over Na₂SO₄, filtered, concentrated and trituratedwith pentane to obtain the desired product as an orange solid. (MS,M⁺+H=416.0)

Compound 721 was prepared in an identical manner to Compound 719, usingthe analogous methyl ester starting material.

Preparation of Compound 745

Displacement: 3-Chloromethyl-6-(2-nitro-phenyl)-imidazo[2,1-b]thiazole(0.200 mmol), imidazole (68 mg, 5 eq), and triethylamine (140 ul) inacetonitrile (3 ml) were heated at 110° C. for 30 minutes in amicrowave. The reaction mixture was concentrated to dryness.

Nitro reduction: The above residue was dissolved in methanol (6 ml), anda mixture of NaHS (67 mg, 6 eq) in water (1 ml) was added, and thereaction was stirred at 60° C. overnight. The following morning anotherportion of NaHS (67 mg, 6 eq) was charged and the reaction was heated to85° C. for 3 hours. The reaction was cooled, concentrated to dryness,diluted with CH₂Cl₂ and water and extracted with CH₂Cl₂ (2×40 ml). Thecombined organic layers were dried (Na₂SO₄), and concentrated.

Amide Formation The amine from above, was dissolved in pyridine (3 ml),and stirred with 2-quinoxalyl chloride (46 mg, 1.2 eq) at roomtemperature for 3 hours. The reaction mixture was concentrated todryness and purified by reverse phase prep-HPLC. And lyophilized withHCl to obtain the HCl salt. (MS, M⁺+H=452.1)

Example 2 Identification of Sirtuin Modulators

A fluorescence polarization or mass spectrometry based assay was used toidentify modulators of SIRT1 activity. The same assay may be used toidentify modulators of any sirtuin protein. The fluorescencepolarization assays utilizes one of two different peptides based on afragment of p53, a known sirtuin deacetylation target. Compounds 1-18were tested using a substrate containing peptide 1 having 14 amino acidresidues as follows: GQSTSSHSK(Ac)NleSTEG (SEQ ID NO: 1) wherein K(Ac)is an acetylated lysine residue and Nle is a norleucine. The peptide islabeled with the fluorophore MR121 (excitation 635 nm/emission 680 nm)at the C-terminus and biotin at the N-terminus. The sequence of thepeptide substrate is based on p53 with several modifications. Inparticular, all arginine and leucine residues other than the acetylatedlysine have replaced with serine so that the peptide is not susceptibleto trypsin cleavage in the absence of deacetylation. In addition, themethionine residue naturally present in the sequence has been replacedwith the norleucine because the methionine may be susceptible tooxidation during synthesis and purification. Compounds 19-56 were testedusing a substrate containing peptide 2 having 20 amino acid residues asfollows: EE-K(biotin)-GQSTSSHSK(Ac)NleSTEG-K(MR21)-EE-NH₂ (SEQ ID NO: 2)wherein K(biotin) is a biotinolated lysine residue, K(Ac) is anacetylated lysine residue, Nle is norleucine and K(MR121) is a lysineresidue modified by an MR121 fluorophore. This peptide is labeled withthe fluorophore MR121 (excitation 635 nm/emission 680 nm) at theC-termini and biotin at the N-termini. The sequence of the peptidesubstrates are based on p53 with several modifications. In particular,all arginine and leucine residues other than the acetylated lysineresidues have replaced with serine so that the peptides are notsusceptible to trypsin cleavage in the absence of deacetylation. Inaddition, the methionine residues naturally present in the sequenceshave been replaced with the norleucine because the methionine may besusceptible to oxidation during synthesis and purification. As analternative substrate in the assay, the following peptide 3 has alsobeen used for testing Compounds 19 through 56:Ac-EE-K(biotin)-GQSTSSHSK(Ac)NleSTEG-K(5TMR)-EE-NH₂ (SEQ ID NO: 3)wherein K(Ac) is an acetylated lysine residue and Nle is a norleucine.The peptide is labeled with the fluorophore 5TMR (excitation 540nm/emission 580 nm) at the C-terminus. The sequence of the peptidesubstrate is also based on p53 with several modifications. In addition,the methionine residue naturally present in the sequence was replacedwith the norleucine because the methionine may be susceptible tooxidation during synthesis and purification.

The peptide substrates were exposed to a sirtuin protein in the presenceof NAD⁺ to allow deacetylation of the substrate and render it sensitiveto cleavage by trypsin. Trypsin was then added and the reaction wascarried to completion (i.e., the deacetylated substrate is cleaved)releasing the MR121 or 5TMR fragment. Streptavidin is then added to thereaction where it can bind both the uncleaved substrate (i.e., anyremaining acetylated substrate) and the non-fluorescent portion of thecleaved peptide substrate (i.e., the biotin containing fragment). Thefluorescence polarization signal observed for the full length peptidesubstrates bound to streptavidin was higher than the fluorescencepolarization signal observed for the released MR121 or 5TMR C-terminalfragment. In this way, the fluorescence polarization obtained isinversely proportional to the level of deacetylation (e.g., the signalis inversely proportional to the activity of the sirtuin protein).Results were read on a microplate fluorescence polarization reader(Molecular Devices Spectramax MD) with appropriate excitation andemission filters.

The fluorescence polarization assays using peptide 1 was conducted asfollows: 0.5 μM peptide substrate and 150 μM βNAD⁺ is incubated with 0.1μg/mL of SIRT1 for 60 minutes at 37° C. in a reaction buffer (25 mMTris-acetate pH8, 137 mM Na—Ac, 2.7 mM K—Ac, 1 mM Mg—Ac, 0.05% Tween-20,0.1% Pluronic F127, 10 mM CaCl₂, 5 mM DTT, 0.025% BSA, 0.15 mMNicotinamide). Test compounds 1-18 were solubilized in DMSO and added tothe reaction at 11 concentrations ranging from 0.7 μM to 100 μM.

Fluorescence polarization assays using peptide 2 may be conducted asfollows: 0.5 μM peptide substrate and 120 μM βNAD⁺ were incubated with 3nM SIRT1 for 20 minutes at 25° C. in a reaction buffer (25 mMTris-acetate pH8, 137 mM Na—Ac, 2.7 mM K—Ac, 1 mM Mg—Ac, 0.05% Tween-20,0.1% Pluronic F127, 10 mM CaCl₂, 5 mM DTT, 0.025% BSA). Test compounds19-56 were solubilized in DMSO and added to the reaction at 10concentrations ranging from 300 μM to 0.15 μM in three-fold dilutions.

After the incubation with SIRT1, nicotinamide was added to the reactionto a final concentration of 3 mM to stop the deacetylation reaction and0.5 μg/mL of trypsin was added to cleave the deacetylated substrate. Thereaction was incubated for 30 minutes at 37° C. in the presence of 1 μMstreptavidin. Fluorescent polarization was determined at excitation (650nm) and emissions (680 nm) wavelengths. The level of activity of thesirtuin protein in the presence of the various concentrations of testcompound is then determined and may be compared to the level of activityof the sirtuin protein in the absence of the test compound, and/or thelevel of activity of the sirtuin proteins in the negative control (e.g.,level of inhibition) and positive control (e.g., level of activation)described below.

For the Fluorescence Polarization assays, a control for inhibition ofsirtuin activity is conducted by adding 1 μL of 500 mM nicotinamide as anegative control at the start of the reaction (e.g., permitsdetermination of maximum sirtuin inhibition). A control for activationof sirtuin activity was conducted using 3 nM of sirtuin protein, with 1μL of DMSO in place of compound, to reach baseline deacetylation of thesubstrate (e.g., to determine normalized sirtuin activity).

The mass spectrometry based assay utilizes a peptide having 20 aminoacid residues as follows:Ac-EE-K(biotin)-GQSTSSHSK(Ac)NleSTEG-K(5TMR)-EE-NH2 (SEQ ID NO: 3)wherein K(Ac) is an acetylated lysine residue and Nle is a norleucine.The peptide is labeled with the fluorophore 5TMR (excitation 540nm/emission 580 nm) at the C-terminus. The sequence of the peptidesubstrate is based on p53 with several modifications. In addition, themethionine residue naturally present in the sequence was replaced withthe norleucine because the methionine may be susceptible to oxidationduring synthesis and purification.

The mass spectrometry assay is conducted as follows: 0.5 μM peptidesubstrate and 120 μM βNAD⁺ is incubated with 10 nM SIRT1 for 25 minutesat 25° C. in a reaction buffer (50 mM Tris-acetate pH 8, 137 mM NaCl,2.7 mM KCl, 1 mM MgCl₂, 5 mM-DTT, 0.05% BSA). Test compounds may beadded to the reaction as described above. The SirT1 gene is cloned intoa T7-promoter containing vector and transformed into BL21(DE3). Afterthe 25 minute incubation with SIRT1, 10 μL of 10% formic acid is addedto stop the reaction. Reactions are sealed and frozen for later massspec analysis. Determination of the mass of the substrate peptide allowsfor precise determination of the degree of acetylation (i.e. startingmaterial) as compared to deacetylated peptide (product).

For the mass spectrometry based assay, a control for inhibition ofsirtuin activity is conducted by adding 1 μL of 500 mM nicotinamide as anegative control at the start of the reaction (e.g., permitsdetermination of maximum sirtuin inhibition). A control for activationof sirtuin activity is conducted using 10 nM of sirtuin protein, with 1μL of DMSO in place of compound, to determine the amount ofdeacetylation of the substrate at a given timepoint within the linearrange of the assay. This timepoint is the same as that used for testcompounds and, within the linear range, the endpoint represents a changein velocity.

For each of the above assays, SIRT1 protein was expressed and purifiedas follows. The SirT1 gene was cloned into a T7-promoter containingvector and transformed into BL21(DE3). The protein was expressed byinduction with 1 mM IPTG as an N-terminal His-tag fusion protein at 18°C. overnight and harvested at 30,000×g. Cells were lysed with lysozymein lysis buffer (50 mM Tris-HCl, 2 mM Tris[2-carboxyethyl]phosphine(TCEP), 10 μM ZnCl₂, 200 mM NaCl) and further treated with sonicationfor 10 min for complete lysis. The protein was purified over a Ni-NTAcolumn (Amersham) and fractions containing pure protein were pooled,concentrated and run over a sizing column (Sephadex S200 26/60 global).The peak containing soluble protein was collected and run on anIon-exchange column (MonoQ). Gradient elution (200 mM-500 mM NaCl)yielded pure protein. This protein was concentrated and dialyzed againstdialysis buffer (20 mM Tris-HCl, 2 mM TCEP) overnight. The protein wasaliquoted and frozen at −80° C. until further use.

Sirtuin modulating compounds that activated SIRT1 were identified usingthe assay described above and are shown below in Table 4. Sirtuinmodulating compounds that inhibited SIRT1 were identified using theassay described above and are shown below in Table 5. The ED₅₀ valuesfor the activating compounds in the fluorescence polarization assay (FP)or mass spectrometry assay (MS) are represented by A′ (ED₅₀=<5 μM), A(ED₅₀=5-50 μM), B (ED₅₀=51-100 μM), C (ED₅₀=101-150 μM), and D(ED₅₀=>150 μM). NT means that the compound was not tested using theindicated assay. NA means that the compound was not active in theindicated assay. Fold activation, as determined by MS is represented byA (Fold activation >250%), B (Fold Activation <250%), or C (no foldactivation). The ED₅₀ of resveratrol for activation of SIRT1 is 16 μMand the fold activation of resveratrol for SIRT1 in the MS assay isapproximately 200%. Similarly, the IC₅₀ values for the inhibitingcompounds are represented by A (IC₅₀=<50 μM), B (IC₅₀=51-100 μM), C(IC₅₀=101-150 μM), and D (IC₅₀=>150 μM).

TABLE 4 Sirt1 Activators ED₅₀ ED₅₀ FOLD COMPOUND FP MS ACT. NO [M + H]+STRUCTURE ASSAY ASSAY MS 1

A NT 2

D NT 3

B NT 4

N/A NT 5

B NT 6

D NT 7 346

A NT 8

B NT 9

C NT 10

D NT 19 409.6

D D C 20 401.3

D D C 21 399.1

D D C 22 414.0

D D C 24 359.4

D D C 27 376.1

D D C 29 385.1

D D C 31 360.1

D D C 32 400.0

D D C 33 376.1

D D C 34 406.3

D D C 35 346.5

D D C 36 376.7

D D C 37 316.4

D D C 38 401.0

D D C 39 399.1

D D C 40 414.2

D D C 41 414.2

D D C 42 359.1

A A B 43 359.1

A A B 45 341.0

D D C 46 376.1

D D C 48 406.3

D D C 49 360.1

D A B 50 400.0

NT D 51 360.1

A A B 52 376.1

A A B 53 406.1

D D C 54 346.4

NT D 55 376.1

A A B 56 316.0

D A B 57 407.1

A 58 377.1

NA 59 318.1

NA 60 413.1

A 61 413.1

NA 62 349.0

NA 63 377.1

A 64 360.1

A 65 383.0

NA 66 407

A 67 377

A 68 360

A 69 377.1

A B 70 360.1

A B 71 376.1

A B 72 422.1

NT 73 377

NT 74 412

D C 75 407.1

A B 76 360.1

A B 77 376.1

A B 78 445.1

79 338

D C 80 355

A B 81 354.5

A B 82 402

C 83 355

A B 84 417

A B 85 335.1

D C 86 375.1

D C 87 375.1

D C 88 382.1

D C 89 365.1

D C 90 381.1

91 412.1

D C 92 308.1

A B 93 329.1

A B 94 434.1

A B 95 345.1

A′ B 96 376.1

A′ B 97 359.1

A′ B 98 408.1

D C 99 391

A′ A 100 376

A B 101 376

A A 102 406

A A 103 374

D C 104 346.1

A′ B 105 330.1

A′ B 106 406.1

A′ B 107 488

A B 108 324

NA 109 401

A B 110 381

C 111 359

A B 112 376

A B 113 375

A B 114 392

A′ A 115 422

A A 116 386

C 117 388

A A 118 410

A A 119 375

A B 120 391

NT 121 414

D C 122 417

C 123 474

NT 124 391

A B 125 433

B B 126 374

A B 127 355

A A 128 388

A B 129 418

A′ A 130 358

A B 131 418

A A 132 350

A A 133 480

A′ B 134 466

A′ B 135 452

A′ B 136 434

D C 137 420

D 138 471

A B 139 488

A B 141 374.1

A′ B 142 374.1

A′ A 143 410.1

D 144 427.1

D C 145 397.1

A′ B 146 397.1

A′ B 147 392.1

D 148 405.2

D 149 359.0

A′ B 150 375.0

A′ B 151 375.0

D C 152 392.1

D C 153 490

A B 154 473

C A 155 490

A B 156 433

A B 157 416

D B 158 433

A B 159 474

A B 160 457

B A 161 474

A A 162 392.1

A′ B 163 422.1

A′ B 164 488

A B 165 416

D A 166 457

A B 167 373

A B 168 388.1

NA C 169 343.1

A′ B 174 479

B A 175 323.1

B B 176 354.1

B B 177 324.1

D B 178 437

A A 179 467

A′ A 180 358.0

A B 181 405.0

A′ A 182 359.0

A′ A 183 358.0

A A 184 375.0

A′ B 185 374.9

A′ A 186 405.3

NA C 187 358.9

A B 188 358.9

NA C 189 375.2

NA C 190 375.3

A′ B 191 375.0

A′ B 192 375.0

A B 193 358.0

NA C 194 422.3

NA C 195 375.9

NA C 196 374.9

A B 197 391.1

A′ B 198 422.4

A B 199 375.9

A′ B 200 375.4

A′ B 201 391.9

A B 202 392.4

A′ B 203 380

A′ B 204 410

A′ A 205 437

A A 206 467

A A 207 478

A A 208 508

A A 209 479

A A 210 509

A B 211

A′ B 212 362

A B 213 392

A′ B 214 392

A B 215 392

A′ B 216 362

A′ B 217 422

A′ A 218 405

A′ A 219 419

A A 220 423

NA C 221 321.4

A B 222 366.8

A B 223 337.4

A′ B 225 332.4

NA C 226 412.5

227 333.4

NA C 228 391.5

A A 229 418.5

NA C 230 459.5

NA C 231 425.5

NA C 232 423.5

NA C 234 449.5

NA C 235 436.3

NA C 236 423.5

NA C 237 450.5

NA C 238 480.5

A′ B 239 466.5

A B 240 392.4

NA C 241 397.2

A′ B 244 332.4

NA C 245 423.5

A′ B 246 391.5

A′ B 247 413.5

NA C 248 467.4

A B 249 395.9

NA C 250 385.5

NA C 251 425.5

NA C 252 453.5

NA C 253 373.1

NA C 254 373

A′ B 255 403

A′ B 256 356

NA C 257 413.1

NA C 258 383.1

A B 259 405.1

A′ A 260 375.1

A′ A 261 375.1

A′ A 262 374.1

A′ B 263 404

A B 264 357

A′ B 265 374.1

A B 266 374

A′ A 267 404

A′ B 268 357

A′ B 270 478

A A 271 508

A′ A 272 392

A′ B 273 422

A A 276 375.1

A′ B 280 381.5

282 386.5

NA C 283 451.6

NA C 284 439.5

NA C 285 440.5

NA C 286 390.1

NA C 287 457.6

288 424.5

A B 289 427.5

A B 290 445.5

NA C 292 420.1

D 293 404.1

A′ A 294 374

A′ B 295 252.1

A′ B 296 376

A B 297 376

A B 298 406

A B 299 359

A B 303 437.5

NA 304 336.4

A′ B 305 414.5

NA 306 424.5

A B 307 382.4

A′ A 308 400.3

NA 309 367.4

NA 310 387.5

NA 311 359

A B 313 418.1

A B 314 388.1

A B 315 388.1

NA 317 392

A′ B 318 392

A B 319 422

A′ B 320 375

A′ B 321 375

A′ B 322 392

NA 323 392

A B 324 422

NA 325 375

NA 326 478

A B 327 461

A A 328 418

B A 329 462

A A 330 443

A A 331 335

A B 332 335.1

A B 333 365

A B 334 340.1

A B 335 340

A B 336 10 370

A B 337 443

NA 338 458

A A 339 376

NA 340 376

NA 341 406

A′ B 342 359

NA 343 406

A B 344 375

A′ B 345 376

NA 346 376

NA 347 406

A′ B 348 359

NA 349 375

NA 350 431.1

B B 351 461

A B 359 472.1

NA 362 502

B B 364 472

D A 367 359.1

NA 369 350.8

NA 370 437.0

NA 371 381.1

NA 372 431.1

NA 373 445.0

NA 374 421.1

NA 375 358.2

NA 376 350.0

NA 377

NA 378 412.1

NA 379 380.0

NA 380 429.9

NA 381 444.1

NA 382 420.0

NA 383 515.7

NA 384 487.8

NA 385 397.2

NA 387 366.8

NA 390 412.5

A B 391 333.4

A B 392 424.5

A B 393 400.3

NA 394 457.6

NA 396 389.5

A B 398 460.1

A B 399 424.5

A B 400 392

NA 401 422

A B 402 375

A′ B 403 375

A′ B 404 376

A′ B 405 406

A′ B 406 359

A′ B 407 359

A′ B 408 359

B B 409 422

NA 410 359

NA 411 422

A B 412 406

NA 413 385.1

B B 414 385

A B 415 415

B B 416 368

NA 419 406

NA 420 376

NA 421 406

A B 422 382

A B 423 382

A′ B 424 382

NA 425 412

NA 426 412

A′ B 427 365

A′ B 428 365

NA 429 376

A′ B 430 406

A A 431 359

A B 436 445.1

A A 437 375.1

A B 438 375

A′ A 439 405

A′ B 440 468

A′ A 441 470

A′ A 442 472

A′ A 443 436

A A 444 464

A A 445 432

A B 446 424

A B 447 484

NA 448 510

NA 449 376

NA 450 392

NA 451 376

A B 452 406

A B 453 359

A B 454 376

A B 455 376

A′ B 456 376

A′ B 457 406

A B 458 359

A′ B 459 359

A′ B 460 359.4

A B 461 367.4

NA 462 391.5

A′ B 463 375.4

A B 465 395.9

NA 466 445.5

NA 467 427.2

NA 468 345.0

A B 469 435.4

NA 470 365.0

NA 471 488.9

NA 472 437.5

NA 473 420.5

A′ B 474

NA 475 408.6

NA 476 410.9

NA 477 435.9

NA 478 404.8

NA 479 420.9

NA 481 428.5

A B 482 344.1

A′ B 483 364.1

NA 484 448.6

A B 485 363.5

A A 486 385.9

NA 487 396.1

NA 488 435.1

NA 489 410.1

NA 490 434.9

NA 491 420.5

NA 492 404.0

NA 493

NA 494 422.9

NA 495 366.0

NA 496 349.9

NA 497 346.0

NA 498 406.5

A B 499 362.1

NA 500

NA 501 347.1

NA 502 363.1

NA 503 443.1

A′ A 504 358

A B 505 359

NA 506 429.1

A A 507 388.1

A′ A 508 366.1

A′ A 510 457

A′ A 511 460

NA 512 484

A′ A 513 470

A′ A 514 466

A B 515 398

A′ B 516 398

NA 517 428

NA 518 381

A′ B 519 381

A′ B 520 428

A′ B 521 375.0

A′ B 522 405.0

A A 523 359.0

A B 524 358.0

A′ B 525 375.0

A′ B 526 400.0

A′ A 527 398.0

A′ B 528 412.9

A′ B 529 399.1

A′ B 530 359.0

A′ B 531 345.0

A′ B 532 345.0

A′ B 533 315.0

A′ B 534 358.0

A′ B 535 428.1

A A 536 442.1

A A 537 444.0

A′ A 538 443.1

A A 539 457.1

A′ A 540 402.1

A A 541 443.1

A B 542 444

A A 543 420

A′ A 544 474

A A 545 378.1

A B 546 408

A B 547 369

A′ B 548 370

A′ B 556 365.1

A′ A 557 542.1

NA 558 442.1

A′ A 559 508

NA 560 470

NA 561 382

A′ B 562 382

A′ B 563 412

A B 565 400.8

NA 566 409.9

A B 567 374.9

NA 568 367.0

NA 569 374.9

NA 570 361.0

NA 571 444.0

A B 572 429.9

B B 573 431.8

NA 574 376.2

NA 575 439.9

NA 576 376.1

NA 577 400.1

NA 578 368.1

NA 579 401.1

NA 580 374.0

NA 581 334.0

A B 582 400.0

NA 583 374.1

NA 584 360.0

NA 585 443.1

A B 587 427.1

A′ B 588 520

A A 589 490

A′ A 590 474

A B 591 458

A′ A 592 455

A′ B 593 473

A′ A 594 466

A B 596 467.4

A′ B 597 377.2

A′ B 599 422.3

A′ B 600 422.5

D B 601 405.5

NA 604 360.4

NA 605 377.4

NA 607 415.4

NA 608 332.4

NA 609 439.3

NA 610 418.6

NA 611 465.6

A′ B 612 402.5

NA 614 428.5

NA 615 408.6

NA 616 437.5

617 471.1

A′ A 618 469.1

A′ B 619 455

A′ B 620 493.1

A′ B 621 472

A′ A 622 482

A′ A 623 437

NA C 624 458

A′ A 625 496

A′ A 628 574.1

A′ B 629 429.0

A A 630 403.1

A A 631 445.0

A′ B 632 458.1

A A 633 423.3

NA C 634 364.9

D B 635 421.1

D A 636 359.9

B B 637 459.3

D B 638 445.2

D B 639 378.9

A B 640 368.9

NA C 641 334.9

D B 642 446.2

D B 643 535.1

A B 644 434

A A 645 469

A′ A 646 481

A A 647 473.1

A′ A 648 402.1

A′ B 649 512.2

A′ A 650 570.2

NA 651 512.2

A′ A 655 393.0

NA C 656 393.2

NA C 657 423.1

NA C 658 382.1

NA C 659 509.2

NA C 660 408.1

A B 661 408.2

A′ B 662 378.2

NA C 663 438.0

A B 664 477.8

NA C 665 406.1

NA C 666 391.0

NA C 667 448.0

A A 668 410.0

A B 669 392.0

NA C 670 392.0

A B 671 422.0

NA C 672 415.1

NA C 673

NA C 674

NA C 675 407.1

NA C 676

A′ A 677

A′ A 678

A B 679

A B 680 484.2

A′ A 681 522

A A 682 492

A′ A 683 475

A B 684 460

A B 685 456

A′ B 686 475

A′ A 687 467

NA C 688 483

NA C 689 479

A A 690 483

A′ A 692 469

C B 695 454

A′ A 697 596

NA C 698 502

A′ A 699

A A 700 512.2

A′ A 701 477

A′ A 702 534

A A 703 468

NA C 704 454

NA C 705 387

706 445.1

707 386.1

A′ A 708 494.2

A′ A 709 494.1

NA C 710 494.1

A′ A 711

A B 714

A B 715

A′ B 716

A′ A 717

A B 718 444.1

NA C 719 416

A A 720

NA C 721

A A 722 449

A′ A 723 490

A A 724 482

A′ A 725 505

A′ A 726 491

A′ A 727 458

A′ A 728 466

A A 729 481

A′ A 730 488

A A 731 498

A A 732 497

A′ B 733 484.2

A′ A 735 514.2

A′ A 736 514.2

A′ A 737 500.1

A′ A 738 448.1

A A 739 424.1

A′ A 740 377.1

A′ B 741 498.1

A′ A 742 487.1

A′ B 743 466.1

A A 744 437.1

B A 745 452.1

A′ A

TABLE 5 Sirt1 Inhibitors COM- POUND IC₅₀ FP IC₅₀ MS NO [M + H]+STRUCTURE ASSAY ASSAY 13

A 14

B 15

C 23 359.4

D 25 341.3

B 26 341.4

B 28 401.1

D 30 380.0

D 44 341.0

D 47 385.0

B 291

B 652

B 653

C 654

D

Example 3 Identification of Sirtuin Modulators Using SIRT3

A fluorescence polarization assay was used to identify modulators ofSIRT3 activity. The same assay may be used to identify modulators of anysirtuin protein. The assay utilizes a peptide substrate based on afragment of Histone H4, a known sirtuin deacetylation target. Thesubstrate contains a peptide having 14 amino acid residues as follows:Biotin-GASSHSK(Ac)VLK(MR121) (SEQ ID NO: 4) wherein K(Ac) is anacetylated lysine residue. The peptide is labeled with the fluorophoreMR121 (excitation 635 nm/emission 680 nm) at the C-terminus and biotinat the N-terminus.

The peptide substrate is exposed to a sirtuin protein in the presence ofNAD⁺ to allow deacetylation of the substrate and render it sensitive tocleavage by trypsin. Trypsin is then added and the reaction is carriedto completion (i.e., the deacetylated substrate is cleaved) releasingthe MR121 fragment. Streptavidin is then added to the reaction where itcan bind both the uncleaved substrate (i.e., any remaining acetylatedsubstrate) and the non-fluorescent portion of the cleaved peptidesubstrate (i.e., the biotin containing fragment). The fluorescencepolarization signal observed for the full length peptide substrate boundto streptavidin is higher than the fluorescence polarization signalobserved for the released MR121 C-terminal fragment. Therefore, thefluorescence polarization obtained is inversely proportional to thelevel of deacetylation (e.g., the signal is inversely proportional tothe activity of the sirtuin protein). Results are read on a microplatefluorescence polarization reader (Molecular Devices Spectramax MD) withappropriate excitation and emission filters.

The fluorescence polarization assays may be conducted as follows: 0.5 μMpeptide substrate and 50 μM βNAD⁺ is incubated with 2 nM of SIRT3 for 60minutes at 37° C. in a reaction buffer (25 mM Tris-acetate pH8, 137 mMNa—Ac, 2.7 mM K—Ac, 1 mM Mg—Ac, 0.1% Pluronic F127, 10 mM CaCl₂, 1 mMTCEP, 0.025% BSA). Test compounds are solubilized in DMSO and are addedto the reaction at 11 concentrations ranging from 0.7 μM to 100 μM. TheSIRT3 protein used in the assays corresponded to amino acid residues102-399 of human SIRT3 with an N-terminal His-tag. The protein wasoverexpressed in E. coli and purified on a nickel chelate column usingstandard techniques. After the 60 minute incubation with SIRT3,nicotinamide is added to the reaction to a final concentration of 3 mMto stop the deacetylation reaction and 0.5 μg/mL of trypsin is added tocleave the deacetylated substrate. The reaction is incubated for 30minutes at 37° C. in the presence of 1 mM streptavidin. Fluorescentpolarization is determined at excitation (650 nm) and emissions (680 nm)wavelengths. The level of activity of the sirtuin protein in thepresence of the various concentrations of test compound are thendetermined and may be compared to the level of activity of the sirtuinprotein in the absence of the test compound, and/or the level ofactivity of the sirtuin proteins in the negative control (e.g., level ofinhibition) and positive control (e.g., level of activation) describedbelow.

A control for inhibition of sirtuin activity is conducted by adding 30mM nicotinamide at the start of the reaction (e.g., permitsdetermination of maximum sirtuin inhibition). A control for activationof sirtuin activity is conducted using 0.5 μg/mL of sirtuin protein toreach baseline deacetylation of the substrate (e.g., to determinenormalized sirtuin activity).

Sirtuin modulating compounds that activated SIRT3 were identified usingthe assay described above and are shown below in Table 6. Sirtuinmodulating compounds that inhibited SIRT3 were identified using theassay described above and are shown below in Table 7. The ED₅₀ valuesfor the activating compounds are represented by A (ED₅₀=<50 μM), B(ED₅₀=51-100 μM), C (ED₅₀=101-150 μM), and D (ED₅₀=>150 μM). The ED₅₀ ofresveratrol for activation of SIRT3 is >300 uM. Similarly, the IC₅₀values for the inhibiting compounds are represented by A (IC₅₀=<50 μM),B (IC₅₀=51-100 μM), C (IC₅₀=101-150 μM), and D (IC₅₀=>150 μM).

TABLE 6 COM- POUND NO STRUCTURE ED₅₀ 11

N/A 12

N/A

TABLE 7 COM- POUND NO STRUCTURE IC₅₀ 1

N/A 2

N/A 15

B 16

C 17

B 18

B

Example 4 Cell-based Assays of Sirtuin Activity

Fat mobilization assay. 3T3 L1 cells are plated with 2 ml of 30,000cells/ml in Dulbecco's Modified Eagle Medium (DMEM)/10% newborn calfserum in 24-well plates. Individual wells are then allowed todifferentiate by addition of 100 nM Rosiglitazone. Undifferentiatedcontrol cells are maintained in fresh DMEM/10% newborn calf serumthroughout the duration of the assay. At 48 hours (2 days), adipogenesisis initiated by addition of DMEM/10% fetal calf serum/0.5 mM3-isobutyl-1-methylxanthine (IBMX)/1 μM dexamethasone. At 96 hours (4days), adipogenesis is allowed to progress by removal of the media andadding 2 ml of DMEM/10% fetal calf serum to each well along with either10 μg/mL insulin or 100 nM Rosiglitazone. At 144 hours (6 days) and 192hours (8 days), all wells are changed to DMEM/10% fetal calf serum.

At 240 hours (10 days from the original cell plating), test compounds ata range of concentrations are added to individual wells in triplicatealong with 100 nM Rosiglitazone. Three wells of undifferentiated cellsare maintained in DMEM/10% newborn calf serum and three wells ofdifferentiated control cells are maintained in fresh DMEM/10% newborncalf serum with 100 nM Rosiglitazone. As a positive control for fatmobilization, resveratrol (a SIRT1 activator) is used at concentrationsranging in three fold dilutions from 100 μM to 0.4 μM.

At 312 hours (13 days), the media is removed and cells are washed twicewith PBS. 0.5 mL of Oil Red O solution (supplied in Adipogenesis AssayKit, Cat.#ECM950, Chemicon International, Temecula, Calif.) is added perwell, including wells that have no cells as background control. Platesare incubated for 15 minutes at room temperature, and then the Oil Red Ostaining solution is removed and the wells are washed 3 times with 1 mLwash solution (Adipogenesis Assay Kit). After the last wash is removed,stained plates are visualized, scanned or photographed. Dye is extracted(Adipogenesis Assay Kit) and quantified in a plate reader at 520 nM.Quantitative and visual results are shown in FIG. 16.

Primary dorsal root ganglion (DRG) cell protection assay. Test compoundsare tested in an axon protection assay as described (Araki et al. (2004)Science 305(5686):1010-3). Briefly, mouse DRG explants from E12.5embryos are cultured in the presence of 1 nM nerve growth factor.Non-neuronal cells are removed from the cultures by adding5-fluorouracil to the culture medium. Test compounds are added 12 to 24hours prior to axon transections. Transection of neurites was performedat 10-20 days in vitro (DIV) using an 18-gauge needle to remove theneuronal cell bodies.

Example 5 ATP Cell-Based Assay

This example describes the effect of the SIRT1 activator, resveratrol oncellular ATP levels in NCI-H358 cells. Cellular ATP levels are anindirect measurement of cellular metabolic rates and, by extension,mitochondrial function. As SIRT1 activation has been linked to increasedmitochondrial biogenesis in vivo, this study is designed to determine ifresveratrol increases mitochondrial function, using cellular ATP levelsas the readout. The ATP assay is combined with a cellular viabilityassay so that cellular ATP levels can be normalized to viable cells.Cellular ATP levels were measured using the ATPLite 1Step Kit(PerkinElmer) and cellular viability was measured using the cellpermeable dye, AlamarBlue™.

The Cellular ATP Assay is a multiplexed assay that measures both ATPlevels and viability of a given cell sample. This assay is run in a96-well Assay Plate and data are reported as the [ATP]/viability foreach well in the Assay Plate.

The ATPLite 1 Step™ Kit is a single-step luminescent cell-based assayfor detection of ATP. The kit contains lyophilized substrate mixture,comprised of D-luciferin and the firefly (Photinus pyralis) enzymeluciferase. Additionally, the kit contains a detergent-basedreconstitution buffer that induces the lysis of cellular membranes. Theluciferase in the assay mixture catalyzes a reaction between the freecellular ATP and D-luciferin to produce bioluminescence according to theschematic reaction outlined below. The amount of light produced isproportional to the cellular ATP concentration.

The AlamarBlue™ Assay is a single-step assay that utilizes a soluble,non-toxic, cell permeable dye that is added to cell growth media. Thisdye undergoes electron reduction in viable cells but not dead cells. Thereduced dye product gives a fluorescent signal which can be monitoredwith a fluorescence plate reader (excitation 545 nm and emission 575nm). The amount of fluorescence generated in a given well isproportional to the number of viable cells. The viability signalgenerated by this assay is used to normalize the ATP signal from theATPLite 1Step™ assay results.

Preparation of Test Substance for Cellular ATP Assay: Resveratrol wasweighed and placed in a brown vial. The material was dissolved in 100%vehicle (DMSO) to yield a final concentration of 10 mM (stock solution).The stock solution was serially diluted with 100% DMSO as described inSOP 7.10. The final concentrations of resveratrol in the compound platewere 0.008, 0.023, 0.069, 0.206, 0.617, 1.852, 5.556, 16.667, 50 and 150μM.

The effect of resveratrol on cellular ATP levels in NCI-H358 cells (100μL) was examined using the Cellular ATP Assay as described. Theexperimental design is summarized in FIG. 1. In this assay NCI-H358cells (obtained from the American Tissue Culture Collection, ATCC) wereseeded in 96 well microplates (10⁴ cells/well). The NCI-H358 GrowthCulture Media consists of RPMI 1640 Media supplemented with 10% FBS, 100mg/mL streptomycin, and 100 units/mL penicillin. Three replicate cellmicroplates were treated with 15 μL of 10 concentrations of resveratrol(0.008, 0.023, 0.069, 0.206, 0.617, 1.852, 5.556, 16.667, 50 and 150 μM)or 15 μL vehicle (DMSO; final concentration of 0.5%; 12 replicates perplate). After 48 hours of compound treatment under cell growthconditions, plates were removed from incubator, and 15 μl of AlamarBlue™dye was added to each well. Cell microplates were incubated with dye for2 hours under growth conditions, and fluorescence was subsequentlymeasured using a plate reader. Media containing AlamarBlue™ was removed,and plates were washed in 100 μl of PBS per well. This wash was removed,and 200 μl of 1×ATPLite 1Step reagent was added to each well.Luminescence was then measured using a plate reader. The ATP signal foreach well, measured by the luminescence scan, was normalized to itscorresponding cell viability value, measured by the fluorescence scan,to generate the average ATP level per viable cell unit (ATP/vCell). TheATP/vCell for each treatment was then normalized to the average vehicleATP/vCell for its respective cell microplate, yielding the normalizedATP/vCell (norm. ATP/vCell). Finally, the norm. ATP/vCell for eachunique treatment was averaged across plate replicates, generating theaverage norm. ATP/vCell. Doses of resveratrol that increase cellular ATPlevels have normalized ATP/vCell values greater than 1.0. Theconcentration of resveratrol which gives the 50% of the maximum increasein normalized ATP/vCell (EC50 ATP) was determined by a best-fit curveanalysis using a sigmoidal dose-response curve model.

The ATP levels of cells treated with 10 concentrations of resveratrol orvehicle alone were measured. Each of these ATP levels was normalized tothe cell viability in the corresponding treatment well, generating theATP/vCell value. Each ATP/vCell value was subsequently normalized to itsaverage Vehicle ATP/vCell values for its respective cell microplate.

Data are represented as the normalized ATP/vCell (arbitrary units). FIG.2 shows the best-fit, sigmoidal dose-response curve for the 10concentrations of resveratrol plotted against their correspondingnormalized ATP/vCell values. These values represent an average of threeplate replicates. Resveratrol increases cellular ATP levels in NCI-H358cells in a dose-dependent manner. The maximum increase in cellular ATPlevels was 3.0 fold and occurred with treatment of 50 μM resveratrol.The EC50 ATP for resveratrol was determined to be 29 μM.

Example 6 Screening of Test Compounds in ATP Cell-Based Assay

A number of compounds were screened for their affect on ATP levels inthe assay as described in Example 5. Results are shown in Table 8. TheED₅₀ values for compounds that raised intracellular ATP levels arerepresented by A (ED₅₀=<50 μM), B (ED₅₀=51-100 μM), C (ED₅₀=101-150 μM),and D (ED₅₀=>150 μM). NA means that the compound was not tested usingthe indicated assay. Similarly, the IC₅₀ values for the compounds thatlowered intracellular ATP levels are represented by A (IC₅₀=<50 μM), B(IC₅₀=51-100 μM), C (IC₅₀=101-150 μM), and D (IC₅₀=>150 μM).

TABLE 8 COMPOUND ED₅₀ ATP IC₅₀ ATP NO STRUCTURE ASSAY ASSAY 52

A 42

A 49

A 115

D 79

A 117

B 120

A 121

NA 123

D 85

NA 86

A 87

NA 88

NA 89

A 90

D 91

A 92

B 93

D 94

D 95

NA 97

A 98

NA 99

A 100

A 101

C 102

A 103

NA 104

A 105

A 133

NA 134

NA 135

A 106

A

EQUIVALENTS

The present invention provides among other things sirtuin-activatingcompounds and methods of use thereof. While specific embodiments of thesubject invention have been discussed, the above specification isillustrative and not restrictive. Many variations of the invention willbecome apparent to those skilled in the art upon review of thisspecification. The full scope of the invention should be determined byreference to the claims, along with their full scope of equivalents, andthe specification, along with such variations.

INCORPORATION BY REFERENCE

All publications and patents mentioned herein, including those itemslisted below, are hereby incorporated by reference in their entirety asif each individual publication or patent was specifically andindividually indicated to be incorporated by reference. In case ofconflict, the present application, including any definitions herein,will control.

Also incorporated by reference in their entirety are any polynucleotideand polypeptide sequences which reference an accession numbercorrelating to an entry in a public database, such as those maintainedby The Institute for Genomic Research (IMGR) (www.tigr.org) and/or theNational Center for Biotechnology Information (NCBI)(www.ncbi.nlm.nih.gov).

Also incorporated by reference are the following: PCT Publications WO2005/002672; 2005/002555; and 2004/016726.

1. A compound of the formula:

or a salt thereof.
 2. A composition comprising a compound of claim 1 ora salt thereof.
 3. A pharmaceutical composition comprising a compound ofclaim 1 or a salt thereof and a pharmaceutically acceptable carrier ordiluent.
 4. A packaged pharmaceutical comprising a compound of claim 1or a salt thereof.
 5. A method for ameliorating at least one symptom ofinsulin resistance, metabolic syndrome, diabetes, or for increasinginsulin sensitivity in a subject, comprising administering to a subjectin need thereof a therapeutically effective amount of a compound ofclaim 1 or a salt thereof.
 6. A method for reducing the weight of asubject, comprising administering to a subject in need thereof atherapeutically effective amount of a compound of claim 1 or a saltthereof.
 7. A method for enhancing motor performance or muscleendurance, decreasing fatigue, or increasing recovery from fatigue,comprising administering to a subject in need thereof a therapeuticallyeffective amount of a compound of claim 1 or a salt thereof.
 8. A methodfor ameliorating at least one symptom of a condition wherein motorperformance or muscle endurance is reduced, wherein the condition is oneor more of age-related muscle wasting, and muscle atrophy and/orcachexia associated with burns, bed rest, limb immobilization, or majorthoracic, abdominal, and/or orthopedic surgery, comprising administeringto a subject in need thereof a therapeutically effective amount of acompound of claim 1 or a salt thereof.
 9. A method for ameliorating atleast one symptom of muscle tissue damage associated with hypoxia orischemia, comprising administering to a subject in need thereof atherapeutically effective amount of a compound of claim 1 or a saltthereof.
 10. A method for increasing muscle ATP levels in a subject,comprising administering to a subject in need thereof a therapeuticallyeffective amount of at least one compound of a compound of claim 1 or asalt thereof.